Surface treatment by water-soluble polymers and lipids/liposomes

ABSTRACT

A method of reducing a friction coefficient of a surface is disclosed herein, comprising attaching a water-soluble polymer to the surface, and contacting the water-soluble polymer with liposomes, thereby coating the surface with an amphiphilic lipid. Further disclosed herein are solutions comprising a water-soluble polymer attachable to the surface, liposomes, and an aqueous carrier, for reducing a friction coefficient of a surface, and methods utilizing same. Articles of manufacture comprising a substrate coated by a water-soluble polymer which is coated by an amphiphilic lipid are also described, as are uses and methods for treating a synovial joint disorder associated with increased articular friction.

RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.15/319,005 filed on Dec. 15, 2016, which is a National Phase of PCTPatent Application No. PCT/IL2015/050606 having International FilingDate of Jun. 15, 2015, which claims the benefit of priority under 35 USC§ 119(e) of U.S. Provisional Patent Application No. 62/012,379 filed onJun. 15, 2014. The contents of the above applications are allincorporated by reference as if fully set forth herein in theirentirety.

FIELD AND BACKGROUND OF THE INVENTION

The present invention, in some embodiments thereof, relates to materialscience and, more particularly, but not exclusively, to methods and/orcompositions for reducing a friction coefficient of a surface of animateor inanimate objects.

Various attempts to provide low friction conditions in aqueous media,and particularly under physiological conditions for treating, interalia, joint dysfunction, have been made.

The major mammalian synovial joints, such as hips and knees, exhibitextremely low levels of friction between the articulating cartilagesurfaces over a range of shear rates from rest to 10⁶ sec⁻¹, up topressures of order 100 atmospheres, a property which no man-madesurfaces can emulate. High friction and corresponding wear of cartilageis a signature of joint pathology [Desrochers et al., Journal of theMechanical Behavior of Biomedical Materials 2013, 25:11-22]. Little isknown of the detailed composition or molecular structure of the veryouter layer of the superficial zone (SZ) of the cartilage tissue,exposed to the synovial cavity. The boundary lubrication of synovialjoints has been attributed to the presence at the surface of hyaluronicacid [Ogston & Stanier, The Journal of Physiology 1953, 119:244-252],lubricin [Raclin et al., Nature 1970, 228:377-378] and aggrecans [Seroret al., Biomacromolecules 2011, 12:3432-3443; Seror et al.,Biomacromolecules 2012, 13: 3823-3832], but these macromolecules, bythemselves or in combination with each other, do not provideparticularly good lubrication at physiological pressures [Seror et al.,Biomacromolecules 2011, 12:3432-3443; Seror et al., Biomacromolecules2012, 13: 3823-3832].

Vecchio et al. [Rheumatology (Oxford) 1999, 38:1020-1021] describe theinjection of dipalmitoylphosphatidylcholine (DPPC) lipid surfactantsolutions in propylene glycol into joints in an attempt to provide atreatment for osteoarthritis.

U.S. Pat. No. 6,800,298 describes a lubricating composition (i.e. alubricant) comprising dextran-based hydrogel with lipids.

U.S. Pat. No. 5,403,592 describes a composition comprising a surfaceactive phospholipid and hyaluronic acid in saline solution as being alubricant suitable for physiological use such as lubrication of joints.

A review by Doughty [Contact Lens and Anterior Eye 1999, 22:116-126]describes various re-wetting, conform, lubricant and moisturizingsolutions and their potential impact on contact lens wearers. Many ofthe solutions described therein include polymers such ashydroxypropylmethylcellulose (HPMC; also known as hypromellose),hydroxyethylcellulose, carboxymethylcellulose, polyethylene glycol,poloxamer, polyvinylpyrrolidone (also known as povidone) and hyaluronicacid (HA).

International Patent Application publication WO 2014/071132 describes acontact lens coupled at its surface to a hyaluronic acid-bindingpeptide, for providing hyaluronic acid to the ocular environment bypretreating the lens with hyaluronic acid and replenishing hyaluronicacid from endogenous or exogenous sources as it is washed away ordegraded.

Liposomes are vesicles whose membranes in most cases are based onphospholipid bilayers. They are generally biocompatible and, whenmodified with other molecules, are widely used in clinical applications,primarily as drug delivery vehicles, as well as in gene therapy and fordiagnostic imaging.

International Patent Application Publication WO 2008/038292 discloses,inter alia, multilamellar vesicles or liposomes (MLVs) of severalphospholipids above their liquid-crystalline-phase to gel-phasetransition temperature (Tm) as possible boundary lubricants in thearticular cartilage environment.

International Patent Application Publication WO 2011/158237 discloses,inter alia, a method for lowering the friction coefficient of surfaces,which is effected by applying gel-phase liposomes onto surfaces to forma boundary lubricant layer, wherein the temperature of the surface atthe time of lubrication is below the phase transition temperature (Tm)of the liposomes. The method is described as being suitable forlubricating biological and non-biological surfaces, including thesurfaces of a biological tissue in a mammalian subject, e.g., fortreating joint dysfunction.

Further studies on surface lubrication by liposomes are described in,for example, Gaisinskaya et al. [Faraday Discuss. 2012, 156:217-233],Goldberg et al. [Advanced Materials 2011, 23:3517-3521], Goldberg et al.[Chemistry and Physics of Lipids 2012, 165:374-381] and Goldberg et al.[Biophys. J. 2011, 100:2403-2411].

The mechanism of hydration lubrication, whereby hydration layers held bysurrounding charges provide effective boundary lubrication even at highpressures, is reviewed by Klein [Friction 2013, 1:1-23].

Additional background art includes U.S. Patent Application PublicationNos. 20040171740, 20060270781, 20100098749 and 20110293699; U.S. Pat.Nos. 7,638,137 and 8,273,366; Benelli [Clinical Ophthalmology 2011,5:783-790]; Brodie et al. [Biomedical Materials 2011, 6:015014]; Davittet al. [Journal of Ocular Pharmacology and Therapeutics 2010,26:347-353]; Di Tizio et al. [Biomaterials, 1998, 19, p. 1877-1884];Itoi et al. [CLAO J. 1995, 21:261-264]; Ludwig & van Ooteghem [J. Pharm.Belg. 1989, 44:391-397]; Mourtas et al. [Langmuir 2009, 25:8480-8488];Kang et al. [Journal of Drug Targeting 2010, 18:637-644]; Lee et al.[PNAS 2006, 103:12999-13003]; Pasquali-Ronchetti [Journal of StructuralBiology 1997, 120:1-10]; Simmons et al. [CLAO J. 2001, 27:192-194];Sorkin et al. [Biomaterials 2103, 34:5465-5475]; Thai et al. [Ophthal.Physiol. Opt. 2002, 22:319-329]; Berry et al. [Hyaluronan in dry eye andcontact lens wearers. In: Lacrimal Gland, Tear Film, and Dry EyeSyndromes 2, D. A. Sullivan, D. A. Dartt and M. A. Meneray, Editors.1998, Plenum Press, NY, pp. 785-790]; and Brochu, Ph.D. Thesis in theUniversité de Sherbrooke, Canada, 2008, Id.: 50177338.

SUMMARY OF THE INVENTION

According to an aspect of some embodiments of the invention, there isprovided a method of reducing a friction coefficient of a surface, themethod comprising contacting the surface with a solution comprising atleast one water-soluble polymer, liposomes, and an aqueous carrier,wherein the water-soluble polymer and the surface are selected such thatthe water-soluble polymer is attachable to the surface.

According to an aspect of some embodiments of the invention, there isprovided a method of reducing a friction coefficient of a surface, themethod comprising attaching at least one water-soluble polymer to thesurface, and contacting the at least one water-soluble polymer withliposomes, thereby effecting coating of the surface by an amphiphiliclipid of the liposomes.

According to an aspect of some embodiments of the invention, there isprovided an article of manufacture comprising a composition-of-matter,the composition-of-matter comprising a substrate coated, on at least aportion of a surface thereof, by at least one water-soluble polymer, theat least one water-soluble polymer being coated by an amphiphilic lipidcomprising at least one charged group, wherein at least a portion ofmolecules of the amphiphilic lipid are oriented such that charged groupsthereof face outwards at a surface of the composition-of-matter.

According to an aspect of some embodiments of the invention, there isprovided an article of manufacture comprising a composition-of-matter,the composition-of-matter comprising a substrate coated, on at least aportion of a surface thereof, by at least one water-soluble polymer, thearticle of manufacture being identified for use in efficiently attachingthereto an amphiphilic lipid so as to reduce a friction coefficient ofthe substrate.

According to an aspect of some embodiments of the invention, there isprovided a solution for reducing a friction coefficient of a surfaceaccording to a method described herein, the solution comprising the atleast one water-soluble polymer, the liposomes, and the aqueous carrier.

According to an aspect of some embodiments of the invention, there isprovided a use of a solution described herein in the manufacture of amedicament for treating a synovial joint disorder associated with anincreased friction coefficient of an articular surface in the synovialjoint.

According to some embodiments of the invention, a molar percentage ofphosphatidylcholine in the liposomes is at least 50%.

According to some embodiments of the invention, a concentration ofphospholipids of the liposomes in the solution is in a range of from 0.5mM to 500 mM. According to some embodiments of the invention, theliposomes are selected from the group consisting of small unilamellarvesicles, large unilamellar vesicles and multilamellar vesicles.

According to some embodiments of the invention, the liposomes comprisemultilamellar vesicles.

According to some embodiments of the invention, the liposomes comprisesmall unilamellar vesicles.

According to some embodiments of the invention, the method furthercomprises modifying the surface so as to obtain a modified surface,wherein the water-soluble polymer and the modified surface are selectedsuch that at least one of the at least one water-soluble polymer isattachable to the modified surface.

According to some embodiments of the invention, the surface is aphysiological surface, and the carrier is a physiologically acceptablecarrier.

According to some embodiments of the invention, the surface is anarticular surface of a synovial joint.

According to some embodiments of the invention, contacting the surfacewith the solution comprises injecting the solution into a synovialcavity.

According to some embodiments of the invention, the method is for use inthe treatment of a synovial joint disorder associated with an increasedfriction coefficient of an articular surface in the synovial joint.

According to some embodiments of the invention, the solution describedherein is for use in the treatment of a synovial joint disorderassociated with an increased friction coefficient of an articularsurface in the synovial joint.

According to some embodiments of the invention, attaching at least onewater-soluble polymer to the surface comprises modifying the surface toobtain a modified surface, wherein the water-soluble polymer is selectedto be attachable to the modified surface.

According to some embodiments of the invention, the at least onewater-soluble polymer comprises a modified water-soluble polymer whichfurther comprises at least one functional group for covalently attachingthe polymer to the surface.

According to some embodiments of the invention, the modifiedwater-soluble polymer comprises at least one functional group forcovalently attaching to the surface.

According to some embodiments of the invention, the functional groupcomprises a dihydroxyphenyl group.

According to some embodiments of the invention, the modifiedwater-soluble polymer is hyaluronic acid conjugated to at least onedopamine moiety via an amide bond.

According to some embodiments of the invention, the surface comprisesamine groups.

According to some embodiments of the invention, the at least onewater-soluble polymer comprises a non-ionic polymer.

According to some embodiments of the invention, the non-ionic polymer isselected from the group consisting of a polyvinylpyrrolidone and apolyethylene glycol.

According to some embodiments of the invention, the at least onewater-soluble polymer comprises an ionic polymer.

According to some embodiments of the invention, the ionic polymer hasfrom 1 to 6 charged groups per 1 kDa.

According to some embodiments of the invention, the ionic polymer is ananionic polymer.

According to some embodiments of the invention, the at least onewater-soluble polymer comprises a biopolymer.

According to some embodiments of the invention, the biopolymer isselected from the group consisting of a mucin, a lubricin and apolysaccharide.

According to some embodiments of the invention, the polysaccharide ishyaluronic acid.

According to some embodiments of the invention, a concentration of thehyaluronic acid is in a range of from 0.01 to 10 mg/ml.

According to some embodiments of the invention, a concentration of thehyaluronic acid is less than 1 mg/ml.

According to some embodiments of the invention, the at least onewater-soluble polymer is selected to enhance an affinity of theliposomes to the surface.

According to some embodiments of the invention, attaching the hyaluronicacid to the surface comprises contacting the surface with a solutioncomprising the hyaluronic acid at a concentration in a range of from0.01 to 10 mg/ml.

According to some embodiments of the invention, the liposomes arecharacterized by a phase transition melting point (Tm) above 37° C.

According to some embodiments of the invention, attaching at least onewater-soluble polymer to the surface is effected by injecting an aqueoussolution of the at least one water-soluble polymer into a synovialcavity.

According to some embodiments of the invention, contacting the at leastone water-soluble polymer with liposomes is effected by injecting anaqueous solution of the liposomes into a synovial cavity comprising theat least one water-soluble polymer.

According to some embodiments of the invention, at least a portion ofthe amphiphilic lipid is in a form of a bilayer, the bilayer having alipophilic region between two hydrophilic regions which comprise chargedgroups.

According to some embodiments of the invention, the bilayer is bound tothe water-soluble polymer by electrostatic attraction.

According to some embodiments of the invention, the synovial jointdisorder is selected from the group consisting of arthritis, traumaticjoint injury, locked joint, and joint injury associated with surgery.

According to some embodiments of the invention, the arthritis isselected from the group consisting of osteoarthritis, rheumatoidarthritis and psoriatic arthritis.

Unless otherwise defined, all technical and/or scientific terms usedherein have the same meaning as commonly understood by one of ordinaryskill in the art to which the invention pertains. Although methods andmaterials similar or equivalent to those described herein can be used inthe practice or testing of embodiments of the invention, exemplarymethods and/or materials are described below. In case of conflict, thepatent specification, including definitions, will control. In addition,the materials, methods, and examples are illustrative only and are notintended to be necessarily limiting.

BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS

Some embodiments of the invention are herein described, by way ofexample only, with reference to the accompanying drawings. With specificreference now to the drawings in detail, it is stressed that theparticulars shown are by way of example and for purposes of illustrativediscussion of embodiments of the invention. In this regard, thedescription taken with the drawings makes apparent to those skilled inthe art how embodiments of the invention may be practiced.

In the drawings:

FIGS. 1A-1B present photographs of a cornea-mimicking lens holder (FIG.1A) and the same holder with a soft contact lens mounted in place (FIG.1B), used in some of the experiments employing a tribometer described inthe Examples section hereinunder, in which the soft contact lens has anexemplary hydrogel surface.

FIG. 2 presents bar graphs showing the friction coefficient of EtafilconA contact lens upon immersion in saline, HA 1 MDa 0.2 mg/ml, MLV HSPCliposomes (45 mM), MLV HSPC liposomes+HA, MLV DMPC (45 mM), or MLVDMPC+HA, followed by rinsing with saline, as measured at a load of 5, 10and 40 grams (corresponding respectively to mean pressures of 0.14, 0.17and 0.27 atmospheres).

FIG. 3 presents bar graphs showing the friction coefficient ofNarafilcon A contact lens upon immersion in saline, HA 1 MDa 0.2 mg/ml,MLV HSPC liposomes (45 mM), MLV HSPC liposomes+HA, MLV DMPC (45 mM), orMLV DMPC+HA, followed by rinsing with saline, as measured at a load of5, 10 and 40 grams (corresponding respectively to mean pressures of0.23, 0.29 and 0.46 atmosphere).

FIG. 4 presents bar graphs showing the friction coefficient of EtafilconA contact lens upon immersion in PBS, solutions of HA, PVP or PEO (0.2mg/ml), a solution of SUV DMPC liposomes (10 mM), or solutions of SUVDMPC liposomes with HA, PVP or PEO, followed by rinsing with PBS, asmeasured at a load of 3 and 10 grams (corresponding respectively to meanpressures of 0.1 and 0.16 atmospheres).

FIG. 5 presents bar graphs showing the friction coefficient of EtafilconA contact lens upon immersion in PBS, solutions of HA or PVP (0.2mg/ml), a solution of SUV HSPC liposomes (10 mM), or solutions of SUVHSPC liposomes with HA or PVP, followed by rinsing with PBS, as measuredat a load of 3 and 10 grams (corresponding respectively to meanpressures of 0.1 and 0.16 atmospheres).

FIG. 6 presents bar graphs showing the friction coefficient ofNarafilcon A contact lens upon immersion in PBS, solutions of HA or PVP(0.2 mg/ml), a solution of SUV DMPC liposomes (10 mM), or solutions ofSUV DMPC liposomes with HA or PVP, followed by rinsing with PBS, asmeasured at a load of 3 and 10 grams (corresponding respectively to meanpressures of 0.18 and 0.26 atmospheres).

FIG. 7 presents bar graphs showing the friction coefficient ofNarafilcon A contact lens upon immersion in PBS, solutions of HA, PVP orPEO (0.2 mg/ml), a solution of SUV HSPC liposomes (10 mM), or solutionsof SUV HSPC liposomes with HA, PVP or PEO, followed by rinsing with PBS,as measured at a load of 3 and 10 grams (corresponding respectively tomean pressures of 0.18 and 0.26 atmospheres).

FIG. 8 (Background art) presents a schematic illustration of the mainmacromolecules at the outer cartilage surface: hyaluronan (darker, thickcurves, blue online), bottle-brush-like aggrecans (red online) andlubricins (lighter, thick curves, green online), adapted from Klein, J.(2009) Science 323, 47-48.

FIGS. 9A-9C present AFM micrograph under water of a mica surface bearingDPPC liposomes that have been mixed with HA for 48 hours at T higherthan Tm (FIG. 9A), compared to AFM micrograph of a mica surface bearingDPPC liposomes that have been mixed for 48 hours at T higher than Tmwithout HA (inset (i)), and a Background art cryo-SEM image of a micasurface bearing DPPC liposomes, adapted from Sorkin et al. [Biomaterials2013, 34:5465-5475] (inset (ii)); AFM micrograph of a mica surfacebearing avidin-bHA-DPPC layers (FIG. 9B), with an inset showing anintact liposome on the same scale for comparison; and a schematicillustration of HA-DPPC complexes formed on top of the avidin layer on asurface (FIG. 9C), drawn based on the AFM micrograph of FIG. 9B.

FIGS. 10A-10C present comparative plots showing normal interaction as afunction of surface separation D between two avidin-bHA-DPPC-bearingsurfaces, measured using a surface force balance (SFB), with fullsymbols denoting first approaches, crossed symbols denoting second orthird approaches and empty symbols denoting receding profiles, and withthe black symbols ring to measurements in pure water, and red symbolsreferring to measurements in 0.15 M KNO₃ salt solution (FIG. 10A); and aclose-up of first approaches profiles that present a rapid decrease inthe surface separation (a kink circled in red in FIG. 10B), at D around100 nm, with the dashed line added as a guide to the eye; and aschematic illustration of the surface force balance (SFB) technique(FIG. 10C), with Kn and Ks being the normal and shear springsrespectively, and D being the surface separation.

FIGS. 11A-11C present graphs showing shear force (Fs) vs. time traces,taken directly from SFB measurements, when two avidin-bHA-DPPC bearingsurfaces slide past against each other in pure water (FIG. 11A), withthe two top traces representing two different amplitudes of back andforth shear motion applied to the upper mica surface, and all the othertraces are the shear responses transmitted to the lateral springs atdifferent surface separations and different pressures; a graph showingshear force as a function of shear velocity at pressure P=161 Atm, asmeasured using a surface force balance (SFB), for the twoavidin-bHA-DPPC bearing surfaces slide past against each other in purewater (FIG. 11B), and a graph showing shear force as a function ofsliding time for a given pressure P=61 Atm and shear velocity v_(s) ofabout 0.4 μm/sec, as measured from the USB for the two avidin-bHA-DPPCbearing surfaces slide past against each other in pure water (FIG. 11C).

FIG. 12 presents shear force (Fs) vs. time traces, taken directly fromSFB measurements, when two avidin+bHA+DPPC bearing surfaces slide pastagainst each other in 0.15M KNO₃ salt solution, with the three traceshaving a zigzag pattern representing three different amplitudes of shearmotion applied to the upper mica surface, and the traces below each ofthe aforementioned three traces representing the corresponding shearresponses transmitted to the lateral springs at different surfaceseparations and different pressures.

FIGS. 13A-13B present graphs showing shear forces (Fs) as a function ofnormal forces (Fn), when two avidin-bHA-DPPC bearing surfaces slide pastagainst each other, across water (black symbols) and across 0.15 M KNO₃salt solution (red symbols), with the shaded area including all the Fsvs. Fn profiles for the avidin-bHA-DPPC-bearing surfaces interactingacross water, and the two Fs vs. Fn profiles refer to the measurementshaving the maximum and the minimum value of effective frictioncoefficient μ at high pressure across water (FIG. 13A), and comparativegraphs showing shear forces (Fs) as a function of normal forces (Fn),when two avidin-bHA-DPPC (dashed black line) and two avidin-bHA (bluesymbols) bearing surfaces slide past against each other, across water(crosses represent data from Seror et al., Biomacromolecules 2012, 13:3823-3832, stars represent data not previously published).

FIGS. 14A-14E present photographs showing extraction of a tendon andassociated sheath (FIG. 14B) from a chicken foot (FIG. 14A), theextracted tendon and sheath (FIG. 14C), cutting of the tendon to allowfree gliding of the tendon in the sheath (FIG. 14D), and the cut tendonand sheath after placement in a tribometer (FIG. 14E).

FIG. 15 presents a scheme depicting a tribometer for testing frictionwithin a sample under a load of 40-80 grams, the tribometer including anF_(N), F_(Z) force sensor connected to a component within the sample (aphotograph showing the depicted tribometer is presented in theright-hand panel.

FIG. 16 is a graph showing shear forces (Fs) and friction coefficient μfor a tendon sliding through a sheath, over the course of 500 cycles ofsliding under a normal force (Fn) of 40 grams, upon immersion in PBS orin a solution of hyaluronic acid (HA), hydrogenated soyphosphatidylcholine small unilamellar vesicles (SUV HSPC), andhydrogenated soy phosphatidylcholine small unilamellar vesicles incombination with hyaluronic acid (SUV HSPC/HA) or with hyaluronic acidwith dopamine functional groups (SUV HSPC/HA-Dopa).

FIG. 17 is a graph showing shear forces (Fs) and friction coefficient μfor a tendon sliding through a sheath, over the course of 500 cycles ofsliding under a normal force (Fn) of 80 grams, upon immersion in PBS orin a solution of hyaluronic acid (HA), hydrogenated soyphosphatidylcholine small unilamellar vesicles (SUV HSPC), andhydrogenated soy phosphatidylcholine small unilamellar vesicles incombination with hyaluronic acid (SUV HSPC/HA) or with hyaluronic acidwith dopamine functional groups (SUV HSPC/HA-Dopa).

FIG. 18 presents bar graphs showing friction coefficient μ for a tendonsliding through a sheath after 500 cycles of sliding under a normalforce (Fn) of 40 or 80 grams, upon immersion in PBS or in a solution ofhyaluronic acid (HA), hydrogenated soy phosphatidylcholine smallunilamellar vesicles (SUV HSPC), and hydrogenated soyphosphatidylcholine small unilamellar vesicles in combination withhyaluronic acid (SUV HSPC/HA) or with hyaluronic acid with dopaminefunctional groups (SUV HSPC/HA-Dopa).

FIG. 19 is a bar graph showing fluorescent intensity of the fluorescentdye DiI on a surface of a tendon following immersion in a solution ofDiI-labeled hydrogenated soy phosphatidylcholine (HSPC) liposomes or ina solution of DiI-labeled HSPC liposomes in combination with hyaluronicacid (HSPC+HA) or with hyaluronic acid with dopamine functional groups(HSPC+HA-DN).

FIGS. 20A-20C present fluorescent images showing the fluorescent dye DiIon a surface of a tendon following immersion in a solution ofDiI-labeled hydrogenated soy phosphatidylcholine (FIG. 20A) liposomes orin a solution of DiI-labeled HSPC liposomes in combination withhyaluronic acid (FIG. 20B) or with hyaluronic acid with dopaminefunctional groups (FIG. 20C).

FIG. 21 is a bar graph showing fluorescent intensity of the fluorescentdye DiI for a gelatin-methacrylate hydrogel following immersion in asolution of DiI-labeled hydrogenated soy phosphatidylcholine (HSPC)liposomes or in a solution of DiI-labeled HSPC liposomes in combinationwith hyaluronic acid (HSPC+HA) or with hyaluronic acid with dopaminefunctional groups at a concentration of 4% (HSPC+HA-DN4%) or 18%(HSPC+HA-DN18%) dopamine per hyaluronic acid repeating (disaccharide)unit.

DESCRIPTION OF SPECIFIC EMBODIMENTS OF THE INVENTION

The present invention, in some embodiments thereof, relates to materialscience and, more particularly, but not exclusively, to methods and/orcompositions for reducing a friction coefficient of a surface of animateor inanimate objects.

Before explaining at least one embodiment of the invention in detail, itis to be understood that the invention is not necessarily limited in itsapplication to the details set forth in the following description orexemplified by the Examples. The invention is capable of otherembodiments or of being practiced or carried out in various ways.

In a search for an improved methodology for lubricating interfaces witha surfaces, including interfaces with physiological surfaces, thepresent inventors have studied the effect of a solution containingliposomes, particularly phosphatidylcholine (PC)-containing liposomes,which are known to be biocompatible, in combination with water-solublepolymers such as hyaluronic acid, polyvinylpyrrolidone and polyethyleneoxide, while using different types of hydrogel surfaces, and havesurprisingly uncovered that this combination considerably exceeds thelubrication effect observed in the presence of liposomes alone orwater-soluble polymer alone, resulting in a synergistic effect inreducing the friction coefficient of the treated surface. Thelubrication effect is mediated by boundary lubrication, that is, it doesnot require the presence of the solution between surfaces in order toreduce friction between the surfaces. Rather, contact with the solutionresults in a treated surface, wherein the surface per se ischaracterized by enhanced lubricity.

Referring now to the drawings, FIGS. 2 and 3 show that exposure ofcontact lenses composed of the exemplary hydrogels etafilcon A (FIG. 2 )and narafilcon A (FIG. 3 ) to liposomes and hyaluronic acid (HA)enhances the lubricity of the hydrogel more effectively than doesexposure to liposomes alone or HA alone (as determined using the corneamodel shown in FIGS. 1A-1B). FIGS. 4-7 show that exposure of etafilcon A(FIGS. 4 and 5 ) and narafilcon A (FIGS. 6 and 7 ) hydrogels toliposomes (small unilamellar vesicles) and hyaluronic acid (HA),polyvinylpyrrolidone (PVP) or polyethylene oxide (PEO) enhances thelubricity of the hydrogel surfaces more effectively than does exposureto liposomes alone or HA, PVP or PEO, and that PVP and PEO are typicallyat least as effective as HA at enhancing lubricity in combination withliposomes. FIGS. 4 and 7 show that PEO exhibits particularly strongsynergy with liposomes at enhancing lubricity, whereas PEO alone doesnot enhance lubricity at all and may even reduce lubricity.

This result surprisingly indicates that a hydrogel surface contactedwith water-soluble polymer (such as HA, PVP or PEO) and liposomes is nota mosaic of a surface coated by water-soluble polymer per se and asurface coated by liposomes per se (which would result in a lubricitywhich is intermediate between the lubricity obtained with water-solublepolymer alone and with liposomes alone), but rather, a surface coatedwith water-soluble polymer and liposomes exhibits a physicalcharacteristic which is not present in surfaces coated by water-solublepolymer alone or liposomes alone, indicating synergy between thewater-soluble polymer and liposomes.

FIGS. 2-7 further show that at relatively low pressures dimyristoylphosphatidylcholine liposomes (which are in a liquid phase) are moreeffective at reducing the lubricity than are hydrogenated soyphosphatidylcholine liposomes (which are in a solid phase), whereas athigher pressures, hydrogenated soy phosphatidylcholine liposomes aremore effective.

The present inventors have further studied the effect of phospholipidscomplexed with a water-soluble polymer such as hyaluronic acid which isattached to surfaces, and have uncovered that suchphospholipid-water-soluble polymer complexes form boundary layers whichexhibit an exceptional combination of lubricity and robustness, which isnot exhibited by the water-soluble polymer(s) when used per se. Thepresent inventors have envisioned that such lubricious boundary layersmay be formed on a wide variety of surfaces, including surfaces which donot exhibit affinity to phospholipids per se.

FIGS. 9B and 9C show a surface coated with phosphatidylcholine followingattachment of hyaluronic acid to the surface (by attaching biotinylatedhyaluronic acid to an avidin-coated surface). FIG. 9A shows intactphosphatidylcholine liposomes on a surface, following mixture of theliposomes with hyaluronic acid in solution, but in the absence ofhyaluronic acid attached to the surface.

FIGS. 10A-13B show force measurements between two of the abovementionedsurfaces coated with biotinylated hyaluronic acid andphosphatidylcholine. FIGS. 10A-10B show that the surfaces are separatedby about 22 nm, suggesting that each surface is coated by a layer ofapproximately 11 nm—which corresponds to the combined thickness ofavidin, hyaluronic acid and a phosphatidylcholine bilayer—and that thecoated surfaces are in direct contact with each other. FIG. 11B showsthat the friction has little dependence on sliding velocity, whichindicates a boundary lubrication mechanism. FIGS. 13A-13B show that thefriction coefficient of such surfaces is at an order of magnitude ofonly 10⁻³, even at pressures as high as 220 atmospheres. FIG. 11C showsthat friction between the surfaces remains low during the course of 1hour of continuous shear force application at high pressure, indicatingconsiderable robustness.

FIGS. 16-18 show that hyaluronic acid in combination with liposomes(e.g., small unilamellar vesicles) is effective for reducing frictionassociated with movement of a tendon in manner which is highly robust torepeated cycles of tendon motion, and that functionalization of thehyaluronic acid with dihydroxyphenyl groups (by coupling with dopamine)is even more effective in this respect. FIGS. 19-20C show thatfunctionalization of the hyaluronic acid with dihydroxyphenyl groupsenhances binding of lipids to the tendon surface, suggesting that thereduction of friction corresponds to the degree of lipid binding to thetendon surface which is mediated by the water-soluble polymer (e.g.,unmodified and functionalized hyaluronic acid). FIG. 21 shows thatfunctionalization of the hyaluronic acid with dihydroxyphenyl groupsalso enhances binding of lipids to gelatin methacrylate hydrogel.

These results indicate that treatment of a surface by a combination ofliposomes and attachment of water-soluble polymers (such as HA) resultsin exceptional and robust lubricity. The lubricity does not require anydirect interaction between the liposomes and surface, and does notrequire the water-soluble polymer(s) per se to exhibit a lubricatingeffect.

Without being bound by any particular theory, it is believed that theamphiphilic lipids supplied by the liposomes provide a very low frictioncoefficient as a result of hydration lubrication associated withhydration of the hydrophilic moieties of the lipids. It is furtherbelieved that attachment of water-soluble polymer(s) to a surfaceenhances lubricity by facilitating adherence of the lubricating lipidsto the surface (e.g., anchoring the lipids to the surface), particularlyto a surface which does not normally exhibit affinity to such lipids,thereby enhancing the robustness of the lubricating lipid film, andallowing for enhanced lubricity even under high pressures.

Without being bound by any particular theory, it is further believedthat attachment of the water-soluble polymer(s) to a surface may resultin a smoother surface (e.g., by covering asperities with flexiblepolymer chains thereby further enhancing lubricity.

The exemplified boundary lubrication, which uses molecules native tosynovial joints (e.g., HA and PC lipids) appears to mimic the highlyeffective lubrication in healthy synovial joints, which has hithertobeen unobtainable by prior lubrication techniques in models of synovialjoints. These effects are particularly desirable in the context oftreating synovial joint disorders associated with increased friction ofan articular surface in the afflicted joint, such as arthritis.

Based on the results presented herein, lubrication of a wide variety ofsurfaces besides hydrogels and articular surfaces may be effected, inaccordance with various embodiments of the invention described herein.

Reducing Friction:

As exemplified herein, liposomes and a water-soluble polymer may be usedin combination to reduce a friction coefficient of a variety ofsurfaces.

According to an aspect of some embodiments of the invention, there isprovided a method of reducing a friction coefficient of a surface, themethod utilizing at least one water-soluble polymer (as defined herein)and liposomes (as defined herein).

Any one of the embodiments described herein of any of the aspectsdescribed herein relating to reducing a friction coefficient of asurface may utilize liposomes in accordance with any one of theembodiments described herein with respect to liposomes and/or lipids(e.g., in the section herein relating to liposomes and lipids), as wellas at least one water-soluble polymer in accordance with any one of theembodiments described herein with respect to water-soluble polymers(e.g., in the section herein relating to water-soluble polymers).

In some embodiments, the method comprises attaching the water-solublepolymer(s) to the surface, and contacting the water-soluble polymer(s)with liposomes, thereby effecting coating of the surface by amphiphiliclipids of the liposomes.

In some embodiments, the water-soluble polymer(s) forms a layer whichadheres to the surface as well as to the lipids, thereby mediatingadherence of the lipids to the surface.

In some embodiments, the water-soluble polymer(s) forms a layer whichadheres to the surface at one side of a water-soluble polymer layer andadheres to the lipids at the other side of the water-soluble polymerlayer, thereby mediating adherence of the lipids to the surface.

In some embodiments, attaching at least one water-soluble polymer to thesurface is effected prior to contacting the water-soluble polymer(s)with liposomes.

In some embodiments, attaching at least one water-soluble polymer to thesurface is effected concomitantly and/or subsequent to contacting thewater-soluble polymer(s) with liposomes. In some embodiments, thesurface is contacted with a mixture of the water-soluble polymer(s) andliposomes (e.g., a solution described herein).

Optionally, at least a portion of lipids adhere to the water-solublepolymer(s) prior to attachment of the water-soluble polymer(s) to thesurface. For example, attachment of the water-soluble polymer(s) to thesurface may optionally be effected by a chemical process which is lessrapid than adherence of lipids to the water-soluble polymer(s).

Alternatively or additionally, at least a portion of the water-solublepolymer(s) is attached to the surface prior to adherence of lipids tothe water-soluble polymer(s). For example, attachment of thewater-soluble polymer(s) to the surface may optionally be effected by achemical process which is more rapid than adherence of lipids to thewater-soluble polymer(s).

In some embodiments, the method comprises contacting the surface with aliquid formulation comprising at least one water-soluble polymer(s)(e.g., as described herein in any one of the respective embodiments),liposomes (e.g., as described herein in any one of the respectiveembodiments) and an aqueous carrier (e.g., as described herein in anyone of the respective embodiments). The water-soluble polymer(s) and thesurface are preferably selected such that the water-soluble polymer(s)is attachable to the surface.

Herein throughout, liquid formulations are referred to interchangeablyas “solution”. It is to be noted that the term “solution” encompassesherein throughout any liquid formulation in which the ingredients, e.g.,the water-soluble polymer(s) and/or the liposomes/lipids, are includedwithin a liquid carrier, whereby each of the ingredients can bedissolved or dispersed within the carrier. The term “solution” as usedherein therefore encompasses also “dispersion”, including liquidformulations wherein some ingredients are dissolved and some ingredients(e.g., liposomes) are dispersed. The term “liquid formulation” as usedherein encompasses both a solution and a dispersion.

In some embodiments, attaching the water-soluble polymer(s) to thesurface comprises contacting the surface with a solution comprising awater-soluble polymer at a concentration in a range of from 0.01 to 10mg/ml. In some embodiments, the concentration is in a range of from 0.03to 10 mg/ml. In some embodiments, the concentration is in a range offrom 0.1 to 10 mg/ml. In some embodiments, the concentration is in arange of from 0.3 to 10 mg/ml. In some embodiments, the water-solublepolymer(s) in the solution comprises an ionic polymer (e.g., asdescribed herein in any one of the respective embodiments) at aconcentration described hereinabove, liposomes (e.g., as describedherein in any one of the respective embodiments) and an aqueous carrier(e.g., as described herein in any one of the respective embodiments).

In some embodiments, attaching more than one water-soluble polymer tothe surface comprises contacting the surface with a solution comprisingeach water-soluble polymer at a concentration in a range of from 0.01 to10 mg/ml. In some embodiments, the concentration is in a range of from0.03 to 10 mg/ml. In some embodiments, the concentration is in a rangeof from 0.1 to 10 mg/ml. In some embodiments, the concentration is in arange of from 0.3 to 10 mg/ml. In some embodiments, the water-solublepolymers in the solution comprise at least one ionic polymer (e.g., asdescribed herein in any one of the respective embodiments) at aconcentration described hereinabove, liposomes (e.g., as describedherein in any one of the respective embodiments) and an aqueous carrier(e.g., as described herein in any one of the respective embodiments).

In some embodiments, the water-soluble polymer(s) comprises apolysaccharide (e.g., as described herein in any one of the respectiveembodiments), optionally an ionic polysaccharide, and attaching thepolysaccharide to the surface comprises contacting the surface with asolution comprising the polysaccharide at a concentration in a range offrom 0.01 to 10 mg/ml. In some embodiments, the concentration is in arange of from 0.03 to 10 mg/ml. In some embodiments, the concentrationis in a range of from 0.1 to 10 mg/ml. In some embodiments, theconcentration is in a range of from 0.3 to 10 mg/ml. In someembodiments, the solution is a solution comprising a polysaccharide(e.g., as described herein in any one of the respective embodiments),liposomes (e.g., as described herein in any one of the respectiveembodiments) and an aqueous carrier (e.g., as described herein in anyone of the respective embodiments).

In some embodiments, the water-soluble polymer(s) comprises hyaluronicacid, polyvinylpyrrolidone (PVP) and/or polyethylene oxide (PEO) andattaching the hyaluronic acid, PVP and/or PEO to the surface comprisescontacting the surface with a solution comprising the hyaluronic acid,PVP and/or PEO at a concentration in a range of from 0.01 to 10 mg/ml.In some embodiments, the hyaluronic acid, PVP and/or PEO concentrationis in a range of from 0.03 to 10 mg/ml. In some embodiments, thehyaluronic acid, PVP and/or PEO concentration is in a range of from 0.1to 10 mg/ml. In some embodiments, the hyaluronic acid, PVP and/or PEOconcentration is in a range of from 0.3 to 10 mg/ml. In someembodiments, the solution is a solution comprising hyaluronic acid, PVPand/or PEO (e.g., as described herein), liposomes (e.g., as describedherein in any one of the respective embodiments) and an aqueous carrier(e.g., as described herein in any one of the respective embodiments).

According to another aspect of embodiments of the invention, there isprovided a solution for reducing a friction coefficient of a surfaceaccording to a method described herein, the solution comprising at leastone water-soluble polymer (e.g., as described herein in any one of therespective embodiments), liposomes (e.g., as described herein in any oneof the respective embodiments) and an aqueous carrier (e.g., asdescribed herein in any one of the respective embodiments).

In any of the embodiments described herein, the surface may comprise anytype of material or combination of different types of material,including inorganic material and/or organic material, in crystalline,amorphous and/or gel (e.g., hydrogel) forms, for example, metal,mineral, ceramic, glass, polymer (e.g., synthetic polymer, biopolymer),plant and/or animal biomass, and combinations thereof.

Liposomes and Lipids:

The liposomes and/or lipids according to any one of the embodimentsdescribed in this section may be used in the context of any one of theembodiments of any of the aspects of the inventions described herein.

As used herein and in the art, the term “liposome” refers to anartificially prepared vesicle comprising a bilayer composed of moleculesof an amphiphilic lipid. In an aqueous medium, the bilayer is typicallyconfigured such that hydrophilic moieties of the amphiphilic lipid areexposed to the medium at both surfaces of the bilayer, whereaslipophilic moieties of the lipid are located in the internal portion ofthe bilayer, and therefore less exposed to the medium. Examples ofliposomes which may be used in any one of the embodiments describedherein include, without limitation, small unilamellar vesicles, largeunilamellar vesicles and multilamellar vesicles.

In some embodiments of any one of the embodiments described herein, theliposomes comprise multilamellar vesicles. In some embodiments, theliposomes are primarily (more than 50 weight percents) multilamellarvesicles.

In some embodiments of any one of the embodiments described herein, theliposomes comprise small unilamellar vesicles. In some embodiments, theliposomes are primarily (more than 50 weight percents) small unilamellarvesicles.

In some embodiments of any one of the embodiments described herein, theliposomes comprise large unilamellar vesicles. In some embodiments, theliposomes are primarily (more than 50 weight percents) large unilamellarvesicles.

As used herein, the term “unilamellar” refers to liposomes characterizedby a single lipid bilayer, whereas the term “multilamellar” refers toliposomes characterized by a multiple lipid bilayers, for example,concentric bilayers.

As used herein, the phrase “small unilamellar vesicle” refers tounilamellar liposomes of less than 100 nm in diameter, whereas thephrase “large unilamellar vesicle” refers to unilamellar liposomes atleast 100 nm in diameter.

As used herein, the term “amphiphilic lipid” refers to compoundscomprising at least one hydrophilic moiety and at least one lipophilicmoiety. Examples of amphiphilic lipids include, without limitation,fatty acids (e.g., at least 6 carbon atoms in length) and derivativesthereof such as phospholipids and glycolipids; sterols (e.g.,cholesterol) and steroid acids.

Herein, the term “phospholipid” refers to a compound comprising asubstituted or non-substituted phosphate group and at least one alkylchain (optionally at least two alkyl chains) which is optionally atleast 5 carbon atoms in length, optionally at least 7 atoms in lengthand optionally at least 9 atoms in length. The at least one alkyl chainis optionally a part of an acyl group (e.g., a fatty acid residue) or analkyl group per se (e.g., a fatty alcohol residue). In some embodiments,the phosphate group and on e or two (optionally two) alkyl chains (e.g.,acyl or alkyl) are attached to a glycerol moiety via the oxygen atoms ofglycerol.

In some embodiments of any one of the embodiments described herein, theamphiphilic lipids coating a surface and/or substrate described herein(e.g., a physiological surface, and/or a surface whose frictioncoefficient is being reduced, according to any one of the respectiveembodiments described herein) are in the form of intact liposomes,optionally essentially the same liposomes (e.g., essentially the samemass and molecular composition) contacted with the water-solublepolymer(s).

In some embodiments of any one of the embodiments described herein, atleast a portion of the amphiphilic lipids (optionally substantially allof the lipids) coating the surface are in a form substantially differentthan the liposomes from which the lipids are derived. In someembodiments, during the coating for the surface, liposomes are convertedto open layers (e.g., lipid bilayers and/or lipid monolayers), asopposed to the closed vesicular structure of the liposomes.

Accordingly, any reference herein to coating a surface with liposomesshould not be interpreted as meaning that an obtained coated surfacecomprises liposomes, only that liposomes are utilized by the methodology(e.g., as an ingredient).

As used herein, the term “phospholipid” encompasses lipids having a(phosphorylated) glycerol backbone (e.g., monoacylglyceride and/ordiacylglyceride phospholipids), referred to as glycerophospholipids; andlipids having a (phosphorylated) sphingosine backbone, referred to asphosphosphingolipids (e.g., sphingomyelins).

As used herein, the term “glycolipid” encompasses lipids having a(glycosylated) glycerol backbone (e.g., monoacylglyceride and/ordiacylglyceride glycolipids), referred to as glyceroglycolipids; andlipids having a (glycosylated) sphingosine backbone, referred to asglycosphingolipids (e.g., cerebrosides, gangliosides).

In some embodiments of any one of the embodiments described herein, thehydrophilic moiety is an ionic moiety.

Herein, the phrase “ionic moiety” refers to a moiety which comprises atleast one charged group (as defined herein), and includes anionicmoieties (which have a net negative charge), cationic moieties (whichhave a net positive charge) and zwitterionic moieties (which have anequal number of positive and negative charges, and thus, no net charge).

Without being bound by any particular theory, it is believed that ionicmoieties are particularly effective at binding to water molecules, whichrenders lipid molecules comprising such moieties particularly effectiveat promoting hydration lubrication, in which the bound water moleculesprovide lubrication even at high pressures.

In some embodiments of any one of the embodiments described herein, theamphiphilic lipid comprises at least one phospholipid. Phospholipids aretypically characterized by the presence of an ionic moiety whichincludes a negative charge associated with an oxygen atom in a phosphatemoiety (P—O⁻), although additional charges may be present.

In some embodiments of any one of the embodiments described herein, thephospholipid is a glycerophospholipid. In some embodiments, theglycerophospholipid is a diacylglyceride, comprising two fatty acylgroups and one phosphate group attached to a glycerol backbone.

In some embodiments of any one of the embodiments described herein, aconcentration of phospholipids in liposomes in a solution describedherein is in a range of from 0.5 mM to 500 mM. In some embodiments, theconcentration is in a range of from 1.5 mM to 150 mM. In someembodiments, the concentration is in a range of from 5 mM to 50 mM.

In some embodiments of any one of the embodiments described herein, aconcentration of phospholipids in liposomes in a solution describedherein is in a range of from 0.5 mM to 50 mM. In some embodiments, theconcentration is in a range of from 1.5 mM to 50 mM.

In some embodiments of any one of the embodiments described herein, aconcentration of phospholipids in liposomes in a solution describedherein is in a range of from 5 mM to 500 mM. In some embodiments, theconcentration is in a range of from 5 mM to 150 mM.

In some embodiments of any one of the embodiments described herein, theamphiphilic lipid comprises at least one negatively charged atom and atleast one positively charged atom. In some embodiments, the amphiphiliclipid is zwitterionic, that is, the one or more negative charges in themolecule are balanced out by an equal number of positive charge(s) inthe molecule. In some embodiments, the amphiphilic lipid comprisesexactly one negative charge and one positive charge.

In some embodiments of any one of the embodiments described herein, theamphiphilic lipid comprises at least one phospholipid which comprises aphosphoethanolamine group or N-alkyl derivative thereof.

The phrase “phosphoethanolamine group or N-alkyl derivative thereof”refers to a —O—P(═O)(—O⁻)—OCH₂CH₂NR′R″R′″⁺ group (or a salt thereof),wherein R′, R″ and R′″ are each independently hydrogen or alkyl,preferably C₁₋₄ alkyl. In some embodiments of any one of the embodimentsdescribed herein, the alkyl group(s) attached to the nitrogen atom areeach independently methyl or ethyl. In some embodiments, the alkyl(s) ismethyl. The term “phosphoethanolamine” refers to a group wherein R′, R″and R′″ are each hydrogen. The term “phosphocholine” refers to a groupwherein R′, R″ and R′″ are each methyl.

Without being bound by any particular theory, it is believed that thedistance between the positive and negative charges in aphosphoethanolamine group or N-alkyl derivative thereof is particularlysuitable for binding water molecules and/or promoting hydrationlubrication.

In some embodiments of any one of the embodiments described herein, amolar percentage of the phospholipid described herein (e.g., inliposomes described herein) which comprises a phosphoethanolamine groupor N-alkyl derivative thereof is at least 20%. In some embodiments, themolar percentage is at least 40%. In some embodiments, the molarpercentage is at least 50%. In some embodiments, the molar percentage isat least 60%. In some embodiments, the molar percentage is at least 70%.In some embodiments, the molar percentage is at least 80%. In someembodiments, the molar percentage is at least 90%. In some embodiments,the phospholipid consists essentially of at least one phospholipidcomprising a phosphoethanolamine group or N-alkyl derivative thereof.

In some embodiments of any one of the embodiments described herein, amolar percentage of the amphiphilic lipid described herein (e.g., inliposomes described herein) which consists of at least one phospholipidwhich comprises a phosphoethanolamine group or N-alkyl derivativethereof is at least 20%. In some embodiments, the molar percentage is atleast 40%. In some embodiments, the molar percentage is at least 50%. Insome embodiments, the molar percentage is at least 60%. In someembodiments, the molar percentage is at least 70%. In some embodiments,the molar percentage is at least 80%. In some embodiments, the molarpercentage is at least 90%. In some embodiments, the amphiphilic lipidconsists essentially of at least one phospholipid which comprises aphosphoethanolamine group or N-alkyl derivative thereof.

In some embodiments of any one of the embodiments described herein, theat least one phospholipid comprises at least one phosphatidylcholine.

Herein and in the art, the term “phosphatidylcholine” refers to aglycerophospholipid comprising a phosphocholine group and two fatty acylgroups attached to a glycerol backbone (i.e., a diacylglyceride).

In some embodiments of any one of the embodiments described herein, thephospholipid described herein (e.g., in liposomes described herein) ischaracterized by a molar percentage of phosphatidylcholine (the at leastone phosphatidylcholine described herein) which is at least 20%. In someembodiments, the molar percentage is at least 40%. In some embodiments,the molar percentage is at least 50%. In some embodiments, the molarpercentage is at least 60%. In some embodiments, the molar percentage isat least 70%. In some embodiments, the molar percentage is at least 80%.In some embodiments, the molar percentage is at least 90%. In someembodiments, the phospholipid consists essentially of at least onephosphatidylcholine.

In some embodiments of any one of the embodiments described herein, theamphiphilic lipid described herein (e.g., in liposomes described herein)is characterized by a molar percentage of phosphatidylcholine (the atleast one phosphatidylcholine described herein) which is at least 20%.In some embodiments, the molar percentage is at least 40%. In someembodiments, the molar percentage is at least 50%. In some embodiments,the molar percentage is at least 60%. In some embodiments, the molarpercentage is at least 70%. In some embodiments, the molar percentage isat least 80%. In some embodiments, the molar percentage is at least 90%.In some embodiments, the amphiphilic lipid consists essentially of atleast one phosphatidylcholine.

The fatty acyl groups in a lipid described herein may comprise saturatedfatty acyl groups, monounsaturated fatty acyl groups (having a singleunsaturated bond) and/or polyunsaturated fatty acyl groups (having twoor more unsaturated bonds). In some embodiments, the unsaturated bondsare cis double bonds.

Examples of suitable saturated fatty acyl groups include, withoutlimitation, lauroyl, myristoyl, palmitoyl and stearoyl.

Examples of suitable monounsaturated fatty acyl groups include, withoutlimitation, oleoyl, palmitoleoyl, eicosenoyl, erucoyl, nervonoyl andvaccenoyl.

Examples of suitable polyunsaturated fatty acyl groups include, withoutlimitation, linoleoyl, α-linolenoyl, γ-linolenoyl, dihomo-γ-linolenoyl,stearidonoyl, eicosatetraenoyl, eicosapentaenoyl, docosapentaenoyl,docosahexaenoyl, arachidonoyl and adrenoyl.

In some embodiments of any one of the embodiments described herein, thefatty acyl groups are selected from the group consisting of saturatedand monounsaturated fatty acyl groups. In some embodiments, the fattyacyl groups are saturated fatty acyl groups.

Without being bound by any particular theory, it is believed thatsaturated and monounsaturated fatty acyl groups, particularly saturatedfatty acyl groups, are relatively resistant to chemical reaction such asoxidation, and therefore provide a more resilient system.

In some embodiments of any one of the embodiments described herein, atleast 50% of the fatty acyl groups are the same species of fatty acylgroup (e.g., myristoyl, palmitoyl). In some embodiments, at least 75% ofthe fatty acyl groups are the same species of fatty acyl group. In someembodiments, at least 90% of the fatty acyl groups are the same speciesof fatty acyl group.

Exemplary phospholipids comprising a single species of fatty acyl groupinclude 1,2-dimyristoyl-sn-glyc ero-3-phosphocholine and1,2-dipalmitoyl-sn-glycero-3-phosphocholine.

It is to be appreciated that phase transitions, e.g., melting points(Tm), of the lipid bilayers and liposomes described herein may bedetermined by the skilled person by selecting suitable fatty acyl groupsfor inclusion in the lipids, for example, by selecting relatively shortand/or unsaturated fatty acyl groups (e.g., myristoyl) to obtain arelatively low melting point; and/or by selecting relatively long and/orsaturated fatty acyl groups (e.g., palmitoyl and/or stearoyl) to obtaina relatively high melting point.

In some embodiments of any one of the embodiments described herein, theliposomes described herein are characterized by a phase transitionmelting point above an expected ambient temperature of a surface towhich the liposomes are applied (e.g., as described herein in any one ofthe respective embodiments), such that a surface coated by lipids at theexpected ambient temperature will be coated predominantly by lipids in asolid phase. For example, in some embodiments, liposomes characterizedby a melting point above a physiological temperature (e.g., about 37°C.) are used to coat a physiological surface with lipids (e.g., asdescribed herein in any one of the respective embodiments).

Without being bound by any particular theory, it is believed that lipidcoatings in a solid phase are more resilient against high pressures(e.g., 10 atmospheres or more), and are therefore particularly suitablefor providing lubrication to surfaces (e.g., articular surfaces ofjoints) subject to such high pressures.

In some embodiments of any one of the embodiments described herein, theliposomes described herein are characterized by a phase transitionmelting point below an expected ambient temperature of a surface towhich the liposomes are applied (e.g., as described herein in any one ofthe respective embodiments), such that a surface coated by lipids at theexpected ambient temperature will be coated predominantly by lipids in aliquid phase. For example, in some embodiments, liposomes characterizedby a melting point below a physiological temperature (e.g., about 36°C.) are used to coat a physiological surface with lipids (e.g., asdescribed herein in any one of the respective embodiments).

Without being bound by any particular theory, it is believed that lipidcoatings in a liquid phase provide the most effective lubrication at lowpressures (e.g., below 10 atmospheres), although they may beinsufficiently resilient against higher pressures, and are thereforeparticularly suitable for providing lubrication to surfaces which aregenerally not subjected to such high pressures.

In some embodiments of any one of the embodiments described herein, theliposomes described herein are characterized by a surface charge, whichmay be a positive surface charge or a negative surface charge.

As used herein, the phrase “surface charge” refers to an electric chargeat or near a surface, such as an interface of a liposome with asolution. The phrase “surface charge” encompasses an electric chargeassociated with an electric potential at a surface (e.g., such that apositive electric potential at a surface is indicative of a positivesurface charge, whereas a negative electric potential at a surface isindicative of a negative surface charge); as well as an electric chargewhich is closer to a surface than an electric charge of an opposite sign(e.g., as in a zwitterion wherein the positive charge is closer to thesurface than the negative charge, or vice versa), such that an ion nearthe surface will interact primarily with the electric charge near thesurface (due to the proximity) as opposed to the electric charge of anopposite sign. For example, phosphatidylcholine liposomes typicallyexhibit a positive surface charge because the positive charge of thecholine group is closer to the liposome surface than the negative chargeof the phosphate group.

Optionally, a surface charge of a liposome is associated with a netcharge of the lipid molecules in the liposome, for example, a liposomecomprising anionic lipids has a negative surface charge, and/or aliposome comprising cationic lipids has a positive surface charge.

Alternatively or additionally, a surface charge of a liposome isassociated with a dipole of lipid molecules (e.g., zwitterionic lipidmolecules) in the liposome, for example, a liposome comprising azwitterionic lipid comprising a phosphocholine group may have a positivesurface charge due to the positively charged ammonium groups in thephosphocholine groups being (on average) closer to the surface of theliposomes than the negatively charged phosphate groups in thephosphocholine groups.

The skilled person will be readily capable of determining a surfacecharge. For example, the sign of a surface charge may be determined bycomparing the propensity of a surface (e.g., of a liposome) to bind toanionic vs. cationic compounds (e.g., labeling compounds) and/or by zetapotential measurement (e.g., according to standard techniques used inthe art).

In some embodiments of any one of the embodiments described herein, theliposomes rupture upon contact with the water-soluble polymer(s) (e.g.,on a surface).

Liposome rupture may optionally result in a lipid bilayer in theliposomes being converted from a curved geometry (e.g., as in therelatively spherical liposomes) to a flatter geometry which complementsthe geometry of the surface and/or the water-soluble polymer(s) attachedto the surface (e.g., thereby enhancing affinity of the lipids to thesurface); and/or which results in a flatter, smoother lipid-coatedsurface (e.g., thereby further reducing friction).

Without being bound by any particular theory, it is believed thatrupture of liposomes is induced by affinity of the surface-attachedwater-soluble polymer(s) to the lipids in the liposome, whereby ruptureof the liposomes allows a greater area of the surface-attachedwater-soluble polymer(s) to come into contact with lipids, therebyincreasing an amount of energetically favorable interactions between thewater-soluble polymer(s) and lipid.

In some embodiments of any one of the embodiments described herein,liposomes and water-soluble polymer(s) are selected such that theselected water-soluble polymer(s) is effective at rupturing the selectedliposomes.

Water-Soluble Polymer(s):

The water-soluble polymer(s) according to any one of the embodimentsdescribed in this section may be used in the context of any one of theembodiments of any of the aspects of the inventions described herein,and in combination with liposomes and/or lipids according to any one ofthe embodiments described herein with respect to liposomes and/orlipids.

As used herein, the phrase “water-soluble polymer” encompasses polymershaving a solubility of at least 1 gram per liter in an aqueous (e.g.,water) environment at pH 7 (at 25° C.).

In some embodiments of any of the embodiments described herein, thewater-soluble polymer has a solubility of at least 2 grams per liter(under the abovementioned conditions). In some embodiments, thesolubility is at least 5 grams per liter. In some embodiments, thesolubility is at least 10 grams per liter. In some embodiments, thesolubility is at least 20 grams per liter. In some embodiments, thesolubility is at least 50 grams per liter. In some embodiments, thesolubility is at least 100 grams per liter.

The water-soluble polymer(s) according to any of the embodimentsdescribed herein may comprise at least one ionic polymer and/or at leastone non-ionic polymer which are water-soluble as defined herein.

As used herein, the phrase “non-ionic polymer” refers to a polymer whichdoes not have a charged group.

Examples of suitable non-ionic water-soluble polymers include, withoutlimitation, polyvinylpyrrolidone (also referred to hereininterchangeably as povidone and/or PVP) and polyethylene oxide (alsoreferred to herein interchangeably as PEO, PEG and/or polyethyleneglycol).

As used herein, the phrase “ionic polymer” refers to polymers having atleast one charged group, and encompasses polymers having a net negativecharge (also referred to herein as “anionic polymers”), polymers havinga net positive charge (also referred to herein as “cationic polymers”),and polymers having no net charge (also referred to herein as“zwitterionic polymers”), in an aqueous (e.g., water) environment at pH7.

Herein throughout, the phrase “charged group” refers to any functionalgroup (e.g., a functional group described herein) which is ionic (asdefined herein), including, for example, amine, carboxylic acid,sulfate, sulfonate, phosphate and phosphonate. Thus, each electriccharge in a moiety or molecule is associated with one charged group,although a single charged group (e.g., non-substituted phosphate) may beassociated with more than one electric charge of the same sign (e.g., adianion, a dication).

Herein throughout, the term “ionic” refers to the presence of anelectric charge on at least one atom in a moiety and/or molecule (in atleast 50% of moieties and/or molecules in a population) in an aqueousmedium (e.g., water) at pH 7. The electric charge may be negative(anionic) or positive (cationic). If more than one electric charge ispresent, the electric charges may be negative (anionic) and/or positive(cationic), for example, both a negative and a positive charge may bepresent (zwitterionic).

In some embodiments of any one of the embodiments described hereinrelating to an ionic polymer, at least 75% of the ionic groups in thepolymer have the same charge, that is, at least 75% of the ionic groupsare cationic groups or are anionic groups, such that the polymer issubstantially cationic or anionic, respectively. In some embodiments, atleast 90% of the ionic groups in the polymer have the same charge. Insome embodiments, at least 95% of the ionic groups in the polymer havethe same charge. In some embodiments, at least 98% of the ionic groupsin the polymer have the same charge. In some embodiments, at least 99%of the ionic groups in the polymer have the same charge.

In some embodiments of any one of the embodiments described herein,about 50% of the ionic groups in the polymer have a positive charge andabout 50% of the ionic groups in the polymer have a negative charge,such that the polymer is substantially zwitterionic.

In some embodiments of any one of the embodiments described herein, theionic polymer is characterized by a charge density of from 1 to 6charged groups (ionic groups) per 1 kDa molecular weight of the polymer.In some embodiments, the ionic polymer has from 1.5 to 4 charged groupsper 1 kDa. In some embodiments, the ionic polymer has from 2 to 3charged groups per 1 kDa.

In some embodiments of any one of the embodiments described herein, theionic polymer is characterized by a net charge (i.e., the differencebetween the number of anionic groups and the number of cationic groups)of from 1 to 6 electric charges per 1 kDa molecular weight of thepolymer. In some embodiments, the ionic polymer has a net charge of from1.5 to 4 charges per 1 kDa. In some embodiments, the ionic polymer has anet charge of from 2 to 3 charges per 1 kDa.

In some embodiments of any one of the embodiments described herein, theionic polymer is an anionic polymer, for example, a polymercharacterized by a net negative charge of from 1 to 6 electric chargesper 1 kDa molecular weight of the polymer.

In some embodiments of any one of the embodiments described herein, theionic polymer is a cationic polymer, for example, a polymercharacterized by a net positive charge of from 1 to 6 electric chargesper 1 kDa molecular weight of the polymer.

In some embodiments of any one of the embodiments described herein, theionic polymer is a polysaccharide (which is an ionic polysaccharide).

As used herein throughout, the term “polysaccharide” refers to a polymercomposed primarily (at least 50 weight percent) of monosaccharide unitslinked by glycosidic linkages.

As used herein, the term “monosaccharide” encompasses carbohydrates perse (having the formula Cn(H₂O)n, wherein n is at least 3, typically from3 to 10), as well as derivatives thereof such as amino sugars, in whichat least one hydroxyl group is replaced by an amine or amide group;sugar acids, in which one or two carbon atoms are oxidized to form acarboxylate group; acylated monosaccharides, in which at least onehydroxyl group and/or amine group is substituted by an acyl group (e.g.,acetyl); and sulfated monosaccharides, in which at least one hydroxylgroup is replaced by a sulfate group.

Examples of monosaccharides include, without limitation, hexoses (e.g.,D-hexoses and/or L-hexoses) such as allose, altrose, glucose, mannose,gulose, idose, galactose, talose, psicose, fructose, sorbose andtagatose; pentoses (e.g., D-pentoses and/or L-pentoses) such asarabinose, lyxose, xylose, ribose, ribulose and xylulose; and hexosederivatives such as glucuronic acid, iduronic acid, manuronic acid,guluronic acid, glucosamine and N-alkyl derivatives thereof,galactosamine and N-alkyl derivatives thereof, N-acetylglucosamine,N-acetylgalactosamine, and monosulfated and disulfatedN-acetylgalactosamine, glucuronic acid and iduronic acid.

As used herein, the phrase “glycosidic linkage” refers to a bond betweena hemiacetal group of one compound (e.g., a monosaccharide monomer) anda hydroxyl group of another compound (e.g., another monosaccharidemonomer).

Examples of ionic polysaccharides include, without limitation,hyaluronic acid, chondroitin sulfate, alginic acid, xanthan gum,chitosan and N-alkyl chitosan derivatives.

Hyaluronic acid is an anionic polysaccharide comprising anionicglucuronic acid monomer units along with non-ionic N-acetylglucosaminemonomer units. Hyaluronic acid is an exemplary anionic polymer.

Chondroitin sulfate is an anionic polysaccharide comprising anionicsulfated (e.g., monosulfated and/or disulfated) N-acetylgalactosamine,glucuronic acid and/or iduronic acid monomer units, and anionicglucuronic acid and/or iduronic acid monomer units, along with non-ionicN-acetylgalactosamine monomer units.

Alginic acid is an anionic polysaccharide comprising anionic mannuronicacid and guluronic acid monomer units.

Xanthan gum is an anionic polysaccharide comprising anionic glucuronicacid monomer units, along with non-ionic glucose and mannose monomerunits (including acetyl and/or pyruvyl derivatives thereof).

Chitosan is a cationic polysaccharide comprising cationic glucosaminemonomer units, optionally along with non-ionic N-acetylglucosaminemonomer units. In N-alkyl chitosan derivatives, at least a portion ofthe glucosamine units comprise 1, 2 or 3 alkyl groups, preferably C₁₋₄alkyl, attached to the nitrogen atom. In some embodiments of any one ofthe embodiments described herein, the alkyl groups attached to thenitrogen atoms are each independently methyl or ethyl. In someembodiments, the alkyls are methyl. In some embodiments, the N-alkylatedmonomer unit is N-trimethylglucosamine.

Herein, the terms “hyaluronic acid”, “chondroitin sulfate”, “alginicacid”, “xanthan gum”, “chitosan”, “N-alkyl chitosan derivatives” and anyother ionic compounds named herein, encompass all salts of the namedcompounds along with the non-ionic forms (e.g., acid forms of theanionic polysaccharides, and the free base forms of the cationicpolysaccharides).

Without being bound by any particular theory, it is believed thathyaluronic acid on a surface is particularly effective at binding toliposomes and rupturing them, thereby forming a lipid coating (e.g.,lipid bilayer) with relatively high affinity to the surface.

In some embodiments of any one of the embodiments described herein, thepolysaccharide is in a form of a salt. In some embodiments, the salt isa pharmaceutically acceptable salt (e.g., an ophthalmically acceptablesalt for an ophthalmic application described herein, a salt suitable forparenteral administration for a parenteral application describedherein).

In some embodiments of any one of the embodiments described herein, thepolysaccharide has from 0.2 to 1 charged groups per monosaccharideresidue. In some embodiments, the polysaccharide has from 0.2 to 0.9charged groups per monosaccharide residue. In some embodiments, thepolysaccharide has from 0.3 to 0.7 charged groups per monosaccharideresidue. In some embodiments, the polysaccharide has from 0.4 to 0.6charged groups per monosaccharide residue. In some embodiments, thepolysaccharide has about 0.5 charged groups per monosaccharide residue.It is to be appreciated that a monosaccharide residue may comprise morethan one charged group (e.g., a sulfate group and a carboxylate group).

In some embodiments of any one of the embodiments described herein, themonosaccharide residues comprise no more than one charged group, thatis, 0 or 1 charged group.

In some embodiments of any one of the embodiments described herein, thepolysaccharide is characterized by a net charge (i.e., the differencebetween the number of anionic groups and the number of cationic groups)of from 0.2 to 1 electric charges per monosaccharide residue. In someembodiments, the net charge is from 0.2 to 0.9 electric charges permonosaccharide residue. In some embodiments, the net charge is from 0.3to 0.7 electric charges per monosaccharide residue. In some embodiments,the net charge is from 0.4 to 0.6 electric charges per monosaccharideresidue. In some embodiments, the net charge is about 0.5 electriccharges per monosaccharide residue.

In some embodiments of any one of the embodiments described herein, amolecular weight (i.e., average molecular weight) of the ionic polymeris in a range of from 0.05 to 10 MDa. In some embodiments, the molecularweight is from 0.05 to 5 MDa. In some embodiments, the molecular weightis from 0.5 to 10 MDa. In some embodiments, the molecular weight is from0.5 to 5 MDa. In some embodiments, the ionic polymer is a polysaccharidehaving an aforementioned molecular weight. In some embodiments, theionic polymer is hyaluronic acid having an aforementioned molecularweight.

Herein throughout, an “average molecule weight” of a polymer refers toweight-average molecular weight (Mw).

In some embodiments of any one of the embodiments described herein, thewater-soluble polymer comprises one or more biopolymers.

Herein, the term “biopolymer” refers to a polymer naturally occurring ina living organism. Examples of biopolymers include, without limitation,polynucleotides (e.g., RNA and DNA), polypeptides, polysaccharides andconjugates thereof (e.g., glycoproteins and proteoglycans comprisingpolypeptide and polysaccharide moieties). It is to be appreciated thatbiopolymers may optionally comprise many different species of relatedmonomeric units (e.g., about 20 different types of amino acid residuesand/or various types of monosaccharide moieties) with little or norepetition of the specific species of monomeric units, yet areconsidered polymers because at least some of the monomeric units arerelated in structure (e.g., being amino acid residues or monosaccharidemoieties).

In some embodiments of any one of the embodiments described herein, thebiopolymer(s) comprises a polypeptide (optionally attached to one ormore saccharide moieties) and/or a polysaccharide.

Examples of suitable biopolymers comprising a polypeptide include,without limitation, mucins and lubricin.

Herein, the term “lubricin” refers to a proteoglycan (also known in theart as “proteoglycan 4”) of about 345 kDa. Human lubricin is encoded bythe PRG4 gene. The lubricin optionally comprises a polypeptide sequenceof isoform A and/or isoform B of lubricin, e.g., according to NCBIreference sequence NP_001121180.

Herein, the term “mucin” refers to a family of high molecular weightglycosylated proteins produced by many animals, and encompasses humanmucins such as, for example, mucin 1 (e.g., according to NCBI referencesequence NP_001018016), mucin 2 (e.g., according to NCBI referencesequence NP_002448), mucin 3A (e.g., according to NCBI referencesequence NP_005951), mucin 3B, mucin 4 (e.g., according to NCBIreference sequence NP_004523), mucin SAC, mucin 5B (e.g., according toNCBI reference sequence NP_002449), mucin 6 (e.g., according to NCBIreference sequence NP_005952), mucin 7 (e.g., according to NCBIreference sequence NP_001138478), mucin 8, mucin 12, mucin 13, mucin 15,mucin 16 (e.g., according to NCBI reference sequence NP_078966), mucin17 (e.g., according to NCBI reference sequence NP_001035194), mucin 19,and mucin 20 (e.g., according to NCBI reference sequence NP_001269435).

The polysaccharide may be a non-ionic polymer (as defined herein) or anionic polymer (as defined herein), e.g., according to any of theembodiments described herein relating to an ionic polysaccharide.

Hyaluronic acid (e.g., according to any of the respective embodimentsdescribed herein) is a non-limiting example of a suitable polysaccharideas well as a non-limiting example of a suitable anionic polymer.

In some embodiments of any one of the embodiments described herein, thewater-soluble polymer(s) is selected to enhance an affinity of theliposomes to the surface, that is, the liposome lipids have a greateraffinity to the surface coated by the water-soluble polymer(s) than tothe surface in the absence of the water-soluble polymer(s).

In some embodiments of any one of the embodiments described herein, thewater-soluble polymer(s) comprises an ionic polymer selected such thatthe liposomes are characterized by a surface charge having a signopposite a sign of a net charge of the ionic polymer.

In some embodiments of any one of the embodiments described herein, theliposomes are characterized by a negative surface charge (e.g., asdescribed herein in any one of the respective embodiments) and thewater-soluble polymer(s) comprises an ionic polymer having a netpositive charge (e.g., as described herein in any one of the respectiveembodiments). In some embodiments, the ionic polymer is a polysaccharidehaving a net positive charge (e.g., a cationic polysaccharide describedherein in any one of the respective embodiments).

In some embodiments of any one of the embodiments described herein, theliposomes are characterized by a positive surface charge (e.g., asdescribed herein in any one of the respective embodiments) and thewater-soluble polymer(s) comprises an ionic polymer having a netnegative charge (e.g., as described herein in any one of the respectiveembodiments). In some embodiments, the ionic polymer is a polysaccharidehaving a net negative charge (e.g., an anionic polysaccharide describedherein in any one of the respective embodiments). In some embodiments,the ionic polymer is hyaluronic acid (optionally hyaluronate salts, inaccordance with the definition of “hyaluronic acid” used herein).

In some embodiments of any one of the embodiments described herein, theamphiphilic lipid comprises at least one phospholipid which comprises aphosphoethanolamine group or N-alkyl derivative thereof (e.g., in anyone of the respective embodiments) and the water-soluble polymer(s)comprises an ionic polymer having a net negative charge (e.g., asdescribed herein in any one of the respective embodiments). In someembodiments, the ionic polymer is a polysaccharide having a net negativecharge (e.g., an anionic polysaccharide described herein). In someembodiments, the ionic polymer is hyaluronic acid.

In some embodiments of any of the embodiments described herein, thewater-soluble polymers described herein comprise at least twowater-soluble polymers according to any of the respective embodimentsdescribed herein. In some embodiments, the water-soluble polymerscomprise at least three water-soluble polymers according to any of therespective embodiments described herein.

In some embodiments of any of the embodiments described herein, thewater-soluble polymers described herein comprise at least one biopolymer(according to any of the respective embodiments described herein) incombination with at least one non-ionic polymer (according to any of therespective embodiments described herein). In some embodiments, thewater-soluble polymers described herein comprise at least one mucinand/or lubricin biopolymer (according to any of the respectiveembodiments described herein) in combination with at least one non-ionicpolymer (according to any of the respective embodiments describedherein).

In some embodiments of any of the embodiments described herein, thewater-soluble polymers described herein comprise at least one biopolymer(according to any of the respective embodiments described herein) incombination with at least one ionic polymer (according to any of therespective embodiments described herein). In some embodiments, thewater-soluble polymers described herein comprise at least one mucinand/or lubricin biopolymer (according to any of the respectiveembodiments described herein) in combination with at least one ionicpolymer (according to any of the respective embodiments describedherein).

In some embodiments of any of the embodiments described herein, thewater-soluble polymers described herein comprise at least one ionicpolymer (according to any of the respective embodiments describedherein) in combination with at least one non-ionic polymer (according toany of the respective embodiments described herein).

In some embodiments of any one of the embodiments described herein, amolecular weight (i.e., average molecular weight) of the water-solublepolymer(s) is in a range of from 3 kDa to 10 MDa. In some embodiments,the molecular weight is from 10 kDa to 10 MDa. In some embodiments, themolecular weight is from 20 kDa to 5 MDa. In some embodiments, themolecular weight is from 30 kDa to 2.5 MDa.

In some embodiments of any one of the embodiments described herein, amolecular weight (i.e., average molecular weight) of the water-solublepolymer(s) is in a range of from 3 kDa to 1 MDa. In some embodiments,the molecular weight is from 10 kDa to 1 MDa. In some embodiments, themolecular weight is from 20 kDa to 500 kDa. In some embodiments, themolecular weight is from 30 kDa to 250 kDa. In some embodiments, thewater-soluble polymer(s) comprises a non-ionic polymer (according to anyof the respective embodiments described herein) having an aforementionedmolecular weight. In some embodiments, the non-ionic polymer is PVPand/or PEO having an aforementioned molecular weight.

In some embodiments of any one of the embodiments described herein, amolecular weight (i.e., average molecular weight) of the water-solublepolymer(s) is in a range of from 0.05 to 10 MDa. In some embodiments,the molecular weight is from 0.05 to 5 MDa. In some embodiments, themolecular weight is from 0.5 to 10 MDa. In some embodiments, themolecular weight is from 0.5 to 5 MDa. In some embodiments, thewater-soluble polymer(s) comprises an ionic polymer (according to any ofthe respective embodiments described herein), optionally an ionicpolysaccharide, having an aforementioned molecular weight. In someembodiments, the ionic polymer is hyaluronic acid having anaforementioned molecular weight.

In some embodiments, a concentration of a water-soluble polymer in thesolution (according to any of the respective embodiments describedherein) is in a range of from 0.01 to 10 mg/ml. In some embodiments, theconcentration is in a range of from 0.03 to 10 mg/ml. In someembodiments, the concentration is in a range of from 0.1 to 10 mg/ml. Insome embodiments, the concentration is in a range of from 0.3 to 10mg/ml. In some embodiments, the water-soluble polymer is PVP, PEO and/oran ionic polymer and/or polysaccharide (e.g., as described herein in anyone of the respective embodiments), optionally hyaluronic acid.

In some embodiments, a concentration of each water-soluble polymer inthe solution (according to any of the respective embodiments describedherein) is in a range of from 0.01 to 10 mg/ml. In some embodiments, theconcentration is in a range of from 0.03 to 10 mg/ml. In someembodiments, the concentration is in a range of from 0.1 to 10 mg/ml. Insome embodiments, the concentration is in a range of from 0.3 to 10mg/ml. In some embodiments, the water-soluble polymer is PVP, PEO and/orhyaluronic acid

In some embodiments, a total concentration of water-soluble polymer(s)in the solution (according to any of the respective embodimentsdescribed herein) is in a range of from 0.01 to 20 mg/ml. In someembodiments, the total concentration is in a range of from 0.03 to 20mg/ml. In some embodiments, the total concentration is in a range offrom 0.1 to 10 mg/ml. In some embodiments, the total concentration is ina range of from 0.3 to 10 mg/ml.

In some embodiments, a concentration of a water-soluble polymer in thesolution (according to any of the respective embodiments describedherein) is in a range of from 0.01 to 1 mg/ml. In some embodiments, theconcentration is in a range of from 0.03 to 1 mg/ml. In someembodiments, the concentration is in a range of from 0.1 to 1 mg/ml.

In some embodiments, the concentration is in a range of from 0.3 to 1mg/ml. In some embodiments, the water-soluble polymer is PVP, PEO and/oran ionic polymer and/or polysaccharide (e.g., as described herein in anyone of the respective embodiments), optionally hyaluronic acid.

In some embodiments, a concentration of each water-soluble polymer inthe solution (according to any of the respective embodiments describedherein) is in a range of from 0.01 to 1 mg/ml. In some embodiments, theconcentration is in a range of from 0.03 to 1 mg/ml. In someembodiments, the concentration is in a range of from 0.1 to 1 mg/ml. Insome embodiments, the concentration is in a range of from 0.3 to 1mg/ml. In some embodiments, the water-soluble polymer is PVP, PEO and/orhyaluronic acid

In some embodiments, a total concentration of water-soluble polymer(s)in the solution (according to any of the respective embodimentsdescribed herein) is in a range of from 0.01 to 2 mg/ml. In someembodiments, the total concentration is in a range of from 0.03 to 2mg/ml. In some embodiments, the total concentration is in a range offrom 0.1 to 1 mg/ml. In some embodiments, the total concentration is ina range of from 0.3 to 1 mg/ml.

In some embodiments, a concentration of a water-soluble polymer in thesolution (according to any of the respective embodiments describedherein) is in a range of from 0.01 to 3 mg/ml. In some embodiments, theconcentration is in a range of from 0.01 to 1 mg/ml. In someembodiments, the concentration is in a range of from 0.01 to 0.3 mg/ml.In some embodiments, the concentration is in a range of from 0.01 to 0.1mg/ml. In some embodiments, the water-soluble polymer is PVP, PEO and/orhyaluronic acid

In some embodiments, a concentration of each water-soluble polymer inthe solution (according to any of the respective embodiments describedherein) is in a range of from 0.01 to 3 mg/ml. In some embodiments, theconcentration is in a range of from 0.01 to 1 mg/ml. In someembodiments, the concentration is in a range of from 0.01 to 0.3 mg/ml.In some embodiments, the concentration is in a range of from 0.01 to 0.1mg/ml. In some embodiments, the water-soluble polymer is PVP, PEO and/orhyaluronic acid.

In some embodiments, a total concentration of water-soluble polymer(s)in the solution (according to any of the respective embodimentsdescribed herein) is in a range of from 0.01 to 6 mg/ml. In someembodiments, the total concentration is in a range of from 0.01 to 2mg/ml. In some embodiments, the total concentration is in a range offrom 0.01 to 0.6 mg/ml. In some embodiments, the total concentration isin a range of from 0.01 to 0.2 mg/ml.

In some embodiments of any one of the embodiments described herein, thewater soluble polymer(s) comprises hyaluronic acid, PVP and/or PEO at aconcentration of less than 3 mg/ml. In some embodiments, the hyaluronicacid, PVP and/or PEO concentration is at least 0.01 mg/ml. In someembodiments, the hyaluronic acid, PVP and/or PEO concentration is atleast 0.03 mg/ml. In some embodiments, the hyaluronic acid, PVP and/orPEO concentration is at least 0.1 mg/ml. In some embodiments, thehyaluronic acid, PVP and/or PEO is at least 0.3 mg/ml.

In some embodiments of any one of the embodiments described herein, thewater soluble polymer(s) comprises hyaluronic acid, PVP and/or PEO at aconcentration of less than 0.75 mg/ml. In some embodiments, thehyaluronic acid, PVP and/or PEO concentration is at least 0.01 mg/ml. Insome embodiments, the hyaluronic acid, PVP and/or PEO concentration isat least 0.03 mg/ml. In some embodiments, the hyaluronic acid, PVPand/or PEO concentration is at least 0.1 mg/ml. In some embodiments, thehyaluronic acid, PVP and/or PEO concentration is at least 0.3 mg/ml.

In some embodiments of any one of the embodiments described herein, thewater soluble polymer(s) comprises hyaluronic acid, PVP and/or PEO at aconcentration of less than 0.5 mg/ml. In some embodiments, thehyaluronic acid, PVP and/or PEO concentration is at least 0.01 mg/ml. Insome embodiments, the hyaluronic acid, PVP and/or PEO concentration isat least 0.03 mg/ml. In some embodiments, the hyaluronic acid, PVPand/or PEO concentration is at least 0.1 mg/ml. In some embodiments, thehyaluronic acid, PVP and/or PEO concentration is at least 0.3 mg/ml.

In some embodiments of any one of the embodiments described herein, thewater soluble polymer(s) comprises hyaluronic acid, PVP and/or PEO at aconcentration of less than 0.25 mg/ml. In some embodiments, thehyaluronic acid, PVP and/or PEO acid concentration is at least 0.01mg/ml. In some embodiments, the hyaluronic acid, PVP and/or PEOconcentration is at least 0.03 mg/ml. In some embodiments, thehyaluronic acid, PVP and/or PEO concentration is at least 0.1 mg/ml.

In some embodiments of any one of the embodiments described herein, thewater soluble polymer(s) comprises hyaluronic acid, PVP and/or PEO at aconcentration of less than 0.1 mg/ml. In some embodiments, thehyaluronic acid, PVP and/or PEO concentration is at least 0.01 mg/ml. Insome embodiments, the hyaluronic acid, PVP and/or PEO concentration isat least 0.03 mg/ml.

In some embodiments of any one of the embodiments described herein, aviscosity of the solution (which may reflect at least in part aconcentration of water-soluble polymer(s) therein) is no more than 1000cP (centipoise). In some embodiments, the viscosity is no more than 500cP. In some embodiments, the viscosity is no more than 200 cP. In someembodiments, the viscosity is no more than 100 cP. In some embodiments,the viscosity is no more than 50 cP. In some embodiments, the viscosityis no more than 20 cP. In some embodiments, the viscosity is no morethan 10 cP. In some embodiments, the viscosity is no more than 5 cP. Insome embodiments, the viscosity is no more than 3 cP. In someembodiments, the viscosity is no more than 2 cP. In some embodiments,the solution is an aqueous solution having a viscosity described herein.

Herein, viscosities of a solution are determined at a temperature of 20°C. and at a shear rate of 1 second⁻¹ (unless indicated otherwise).

Attachment of water-soluble polymer to surface:

Attachment of a water-soluble polymer to a surface according to any oneof the embodiments described in this section may be used in the contextof any one of the embodiments of any of the aspects of the inventionsdescribed herein, and in combination with liposomes and/or lipidsaccording to any one of the embodiments described herein with respect toliposomes and/or lipids, and in combination with water-solublepolymer(s) according to any one of the embodiments described herein withrespect to water-soluble polymer(s).

In some embodiments of any one of the embodiments described herein, themethod of reducing a friction coefficient of a surface comprisesmodifying the surface and/or the water-soluble polymer(s), in order tofacilitate attachment of the water-soluble polymer(s) to the surface.The modification may optionally comprise introduction of a functionalgroup or moiety to one material (e.g., the surface or the water-solublepolymer(s)) capable of forming a covalent bond or selective non-covalentbond with the other material (e.g., the water-soluble polymer(s) or thesurface).

In some embodiments of any one of the embodiments described herein, atleast one water-soluble polymer is selected to be attachable to thesurface.

Herein, the phrase “attachable to the surface” and variations thereofrefer to a property of a molecule (e.g., at water-soluble polymerdescribed herein) which renders it capable of attaching via covalent ornon-covalent interactions to the surface. Examples of such interactionsinclude, without limitation, covalent bonds, electrostatic attraction,hydrophobic bonds, hydrogen bonds, and aromatic interactions. It is tobe appreciated that such a property depends on both the properties ofthe molecule (e.g., a water-soluble polymer described herein) and theproperties of the surface, such that a molecule attachable to onesurface is not necessarily attachable to another surface.

In some embodiments of any one of the embodiments described herein, thewater-soluble polymer(s) is attachable to the surface by electrostaticinteractions. In some such embodiments, the water-soluble polymer(s)comprises an ionic polymer having a net charge (e.g., characterized by acharge density described herein) which is of the opposite sign of asurface charge of the surface.

In some embodiments of any one of the embodiments described herein, thewater-soluble polymer(s) is attachable to the surface by covalentbinding and/or by selective non-covalent binding (e.g., as describedherein in any one of the respective embodiments).

In some embodiments of any one of the embodiments described herein, thewater-soluble polymer(s) comprises a modified water-soluble polymer, inwhich a water-soluble polymer (e.g., as described herein in any one ofthe respective embodiments) is modified so as to further comprise atleast one functional group for attaching the polymer to the surface. Insome embodiments, the modified water-soluble polymer comprises at leastone functional group which forms a covalent bond with one or morespecific functional groups (e.g., hydroxy, amine, thiohydroxy and/or oxogroups) which are present on the surface (e.g., a modified surfacedescribed herein or a non-modified surface).

Herein, the phrase “functional group for attaching” encompasses chemicalgroups and moieties of any size and any functionality.

In some embodiments of any one of the embodiments described herein, awater-soluble polymer comprises a dihydroxyphenyl functional group forattaching to a surface.

Herein, the term “dihydroxyphenyl” refers to an aryl group (as definedherein) which is a phenyl substituted by two hydroxyl groups at anypositions thereof. The phenyl may optionally be substituted byadditional substituents (which may optionally comprise additionalhydroxyl groups), to thereby form a substituted dihydroxyphenyl group;or alternatively, the phenyl comprises no substituents other than thetwo hydroxyl groups, such that the dihydroxyphenyl group is anunsubstituted dihydroxyphenyl group.

In some embodiments of any one of the embodiments described herein, thedihydroxyphenyl group is an ortho-dihydroxyphenyl (wherein the hydroxylgroups are attached to the phenyl at adjacent positions) or apara-dihydroxyphenyl (wherein the hydroxyl groups are attached toopposite sides of the phenyl ring), each being a substituted orunsubstituted dihydroxyphenyl. In some such embodiments, theortho-dihydroxyphenyl or para-dihydroxyphenyl is an unsubstituteddihydroxyphenyl.

In some embodiments of any one of the embodiments described herein, thedihydroxyphenyl group is a substituted or unsubstitutedortho-dihydroxyphenyl. In some such embodiments, theortho-dihydroxyphenyl is an unsubstituted ortho-dihydroxyphenyl.

A dihydroxyphenyl group according to any of the respective embodimentsdescribed herein may optionally attach covalently and/or non-covalentlyto a surface according to any one or more attachment mechanism describedfor dihydroxyphenyl (catechol) groups in Lee et al. [PNAS 2006,103:12999-13003] and/or Brodie et al. [Biomedical Materials 2011,6:015014], the contents of each of which are incorporated in theirentirety, and particularly contents regarding binding of dihydroxyphenyl(catechol) groups to surfaces.

Without being bound by any particular theory, it is believed thatortho-dihydroxyphenyl and para-dihydroxyphenyl groups are particularlysuitable for forming covalent bonds by being oxidized (under even verymild oxidizing conditions) to a reactive quinone moiety, which may forcovalent bonds, for example, with amine groups (e.g., primary aminegroups), thiohydroxy groups and other phenyl (e.g., dihydroxyphenyl)groups. It is further believed that ortho-dihydroxyphenyl groups areparticularly suitable for forming non-covalent bonds, for example, withan atom or functional group capable of binding to the two adjacenthydroxyl groups via electrostatic attraction (e.g., upon deprotonationof a hydroxyl group) and/or hydrogen bonds.

In some embodiments of any one of the embodiments described herein, thedihydroxyphenyl group is capable of forming covalent and/or non-covalentbonds with one or more functional groups on a surface, for example,depending on conditions such as pH. For example, a dihydroxyphenyl groupmay optionally be particularly susceptible to covalent bond formationwith an amine group at a relatively basic pH, such as at least about 8.5(e.g., a pH at which the amine is relatively nucleophilic, therebyfacilitating covalent bond formation by nucleophilic attack), whilebeing more susceptible to non-covalent bond formation with an amine at alower pH (e.g., a pH at which the amine is positively charged, therebyfacilitating electrostatic interactions and/or hydrogen bonding).

Modification of a molecule (e.g., water-soluble polymer) withdihydroxyphenyl groups may be performed using any suitable technique forconjugation known in the art. The skilled person will be readily capableof selecting a suitable technique for any given molecule (water-solublepolymer) to be modified.

In some embodiments of any one of the embodiments described herein,modification of a molecule (e.g., water-soluble polymer) is performed byconjugating a compound comprising dihydroxyphenyl group and an aminegroup to a functional group on the molecule being modified which can becoupled to an amine group. Dopamine is a non-limiting example of acompound comprising dihydroxyphenyl group and an amine group. Examplesof functional groups which can be coupled to an amine group include,without limitation, carboxyl groups, which may be coupled (e.g., by acarbodiimide) to an amine to form an amide bond; and aldehyde groups,which may be coupled to an amine to form an imine.

In exemplary embodiments, the modified water-soluble polymer ishyaluronic acid conjugated to at least one dopamine moiety via an amidebond (by conjugation of a dopamine amine group to a hyaluronic acidcarboxylic acid group). A percentage of carboxylic acid groups ofhyaluronic acid conjugated to dopamine may optionally be, for example,in a range of from 0.1% to 90%, optionally from 1% to 50%, optionallyfrom 3% to 25%, and optionally from 10% to 20%.

In some embodiments of any one of the embodiments described herein, thedihydroxyphenyl group is a functional group for attaching (covalentlyand/or non-covalently) to a surface which comprises amine groups,optionally primary amine groups. In some embodiments, such a surfacecomprises proteins, and the amine groups may optionally be lysine sidechain amine groups and/or N-terminal amine groups. In some embodiments,the surface comprises collagen. In some embodiments, the surfacecomprises cartilage (e.g., articular cartilage).

In some embodiments of any one of the embodiments described herein, themethod of reducing a friction coefficient of a surface comprisesmodifying the surface to obtain a modified surface. In some embodiments,the water-soluble polymer(s) is selected to be attachable to themodified surface.

In some embodiments of any one of the embodiments described herein, themodified surface is modified so as to have a functional group whichforms a covalent bond with one or more specific functional groups (e.g.,hydroxy, amine, thiohydroxy and/or oxo groups) and the water-solublepolymer(s) is selected to comprise one or more such groups, therebybeing attachable to the modified surface.

In some embodiments of any one of the embodiments described herein, themodified surface is modified so as to have a moiety capable ofselectively binding (e.g., by non-covalent binding) to a target moiety,and the water-soluble polymer(s) is selected to comprise one or moresuch target moieties, thereby being attachable to the modified surface.In some embodiments, the moiety on the modified surface and the targetmoiety on the water-soluble polymer are each a protein (or a fragmentthereof) and corresponding ligand of the protein (e.g., avidin andbiotin). For example, a protein (or protein domain) may optionally beattached to the surface to form a modified surface, and thewater-soluble polymer(s) is selected to comprise the correspondingligand; or a ligand may optionally be attached to the surface to form amodified surface, and the water-soluble polymer(s) is selected tocomprise a protein (or fragment thereof) which binds to the ligand.

A water-soluble polymer selected to be attachable to the modifiedsurface may be attachable per se, that is, the water-soluble polymer(e.g., as described herein in any one of the respective embodiments) maybe attached to the surface without any modification to the polymer; orthe water-soluble polymer may be a modified water-soluble polymer (e.g.,as described herein in any one of the respective embodiments).

In some embodiments of any one of the embodiments described herein, awater-soluble polymer described herein is a modified water-solublepolymer in which at least a portion of the at least one functional groupfor attaching the polymer to the surface is a target moiety capable ofselective non-covalent binding (e.g., as described herein in any one ofthe respective embodiments). Biotinylated water-soluble polymer is anexample of such a modified water-soluble polymer.

In some embodiments of any one of the embodiments described herein, awater-soluble polymer described herein is selected so as to comprise,without modification to the polymer, a target moiety capable ofselective non-covalent binding (e.g., as described herein in any one ofthe respective embodiments). In some embodiments, the water-solublepolymer is a ligand (e.g., a polysaccharide ligand, a polypeptideligand) and the surface (e.g., modified surface) comprises a protein (orfragment thereof) which binds to such a ligand. In some embodiments, thewater-soluble polymer is a protein (or fragment thereof) and the surface(e.g., modified surface) comprises a ligand which binds to such aprotein (or fragment thereof).

Examples of functional groups for covalent attachment as describedherein, e.g., of a water-soluble polymer (modified or non-modified) to asurface (modified or non-modified), include, without limitation:

-   -   nucleophilic groups such as thiohydroxy, amine (e.g., primary or        secondary amine) and hydroxy, which may form covalent bonds        with, e.g., a functional group comprising a nucleophilic leaving        group, Michael acceptor, acyl halide, isocyanate and/or        isothiocyanate (e.g., as described herein);    -   nucleophilic leaving groups such as halo, azide (—N₃), sulfate,        phosphate, sulfonyl (e.g. mesyl, tosyl), N-hydroxysuccinimide        (NHS) (e.g. NHS esters), sulfo-N-hydroxysuccinimide, and        anhydride, which may form covalent bonds with, e.g., a        nucleophilic group (e.g., as described herein);    -   Michael acceptors such as enones (e.g., maleimide, acrylate,        methacrylate, acrylamide, methacrylamide), nitro groups and        vinyl sulfone, which may form covalent bonds with, e.g., a        nucleophilic group (e.g., as described herein), optionally        thiohydroxy;    -   dihydroxyphenyl groups (according to any of the respective        embodiments described herein, which may form covalent bonds        with, e.g., a nucleophilic group (e.g., as described herein)        and/or a substituted or unsubstituted phenyl group (e.g.,        another dihydroxyphenyl group), as described herein;    -   acyl halide (—C(═O)-halogen), isocyanate (—NCO) and        isothiocyanate (—N═C═S), which may form covalent bonds with,        e.g., a nucleophilic group (e.g., as described herein);    -   carboxylate (—C(═O)OH), which may form covalent bonds with,        e.g., an amine (e.g., primary amine) to form an amide bond; and    -   oxo groups (e.g., aldehydes), which may form covalent imine        bonds with amines (e.g., primary amines).

For any of the abovementioned functional groups for covalent attachment,the functional group may be on the water-soluble polymer (e.g., modifiedwater-soluble polymer) or on the surface (e.g., modified surface).

In some embodiments of any one of the embodiments described herein,attachment of the water-soluble polymer(s) to the surface is effectedvia a linker.

Herein, the term “linker” refers to a compound or moiety which binds(via covalent and/or non-covalent bonds) to two or more substances(e.g., a surface described herein and at least one water-soluble polymerdescribed herein). In embodiments, wherein the linker binds only vianon-covalent bonds, the linker may be regarded as an independentcompound. In embodiments wherein the linker binds to at least onesubstance by at least one covalent bond, the linker may be considered asa moiety which is a part of a substance to which it is bound, forexample, a moiety of a modified surface and/or a modified water-solublepolymer described herein.

In some embodiments of any one of the embodiments described herein, thelinker comprises at least one functional group or moiety which binds tothe surface non-covalently (e.g., as described herein in any one of therespective embodiments), and at least one functional group or moietywhich binds to the water-soluble polymer non-covalently (e.g., asdescribed herein in any one of the respective embodiments). In someembodiments, the water-soluble polymer bound by the functional group ormoiety is a polysaccharide (e.g., as described herein in any one of therespective embodiments), and the linker comprises at least onepolysaccharide-binding polypeptide capable of selectively binding to thepolysaccharide (e.g., as described herein in any one of the respectiveembodiments). In some embodiments, the water-soluble polymer ishyaluronic acid (e.g., as described herein in any one of the respectiveembodiments), and the linker comprises at least one hyaluronicacid-binding polypeptide capable of selectively binding to thehyaluronic acid (e.g., as described herein in any one of the respectiveembodiments).

In some embodiments of any one of the embodiments described herein,attachment of the water-soluble polymer to the surface comprisesattaching the linker to the surface non-covalently, thereby forming amodified surface to which the water-soluble polymer is attachable. Sucha modified surface comprises at least one functional group or moietycapable of binding to the water-soluble polymer non-covalently. In someembodiments, a method described herein comprises attaching the linker tothe surface prior to effecting attachment of the water-soluble polymerto the resulting modified surface.

In some embodiments of any one of the embodiments described herein,attachment of the water-soluble polymer to the surface comprisesattaching the linker to the water-soluble polymer non-covalently,thereby forming a modified water-soluble polymer which is attachable tothe surface. Such a modified water-soluble polymer comprises at leastone functional group or moiety capable of binding to the surfacenon-covalently. In some embodiments, a method described herein comprisesattaching the linker to the water-soluble polymer prior to effectingattachment of the resulting modified water-soluble polymer to thesurface.

In some embodiments of any one of the embodiments described herein, thelinker comprises at least one functional group or moiety which binds tothe surface covalently (e.g., as described herein in any one of therespective embodiments), and at least one functional group or moietywhich binds to the water-soluble polymer covalently (e.g., as describedherein in any one of the respective embodiments).

In some embodiments of any one of the embodiments described herein,attachment of the water-soluble polymer to the surface comprisesattaching the linker to the surface covalently, thereby forming amodified surface to which the water-soluble polymer is attachable. Sucha modified surface comprises at least one functional group or moietycapable of binding to the water-soluble polymer covalently. In someembodiments, a method described herein comprises attaching the linker tothe surface prior to effecting attachment of the water-soluble polymerto the resulting modified surface.

In some embodiments of any one of the embodiments described herein,attachment of the water-soluble polymer to the surface comprisesattaching the linker to the water-soluble polymer covalently, therebyforming a modified water-soluble polymer which is attachable to thesurface. Such a modified water-soluble polymer comprises at least onefunctional group or moiety capable of binding to the surface covalently.In some embodiments, a method described herein comprises attaching thelinker to the water-soluble polymer prior to effecting attachment of theresulting modified water-soluble polymer to the surface.

In some embodiments of any one of the embodiments described herein, thelinker comprises at least one functional group or moiety which binds tothe surface non-covalently (e.g., as described herein in any one of therespective embodiments), and at least one functional group or moietywhich binds to the water-soluble polymer covalently (e.g., as describedherein in any one of the respective embodiments).

In some embodiments of any one of the embodiments described herein,attachment of the water-soluble polymer to the surface comprisesattaching the linker to the surface non-covalently, thereby forming amodified surface to which the water-soluble polymer is attachable. Sucha modified surface comprises at least one functional group or moietycapable of binding to the water-soluble polymer covalently. In someembodiments, a method described herein comprises attaching the linker tothe surface prior to effecting attachment of the water-soluble polymerto the resulting modified surface.

In some embodiments of any one of the embodiments described herein,attachment of the water-soluble polymer to the surface comprisesattaching the linker to the water-soluble polymer covalently, therebyforming a modified water-soluble polymer which is attachable to thesurface. Such a modified water-soluble polymer comprises at least onefunctional group or moiety capable of binding to the surfacenon-covalently. In some embodiments, a method described herein comprisesattaching the linker to the water-soluble polymer prior to effectingattachment of the resulting modified water-soluble polymer to thesurface.

In some embodiments of any one of the embodiments described herein, thelinker comprises at least one functional group or moiety which binds tothe surface covalently (e.g., as described herein in any one of therespective embodiments), and at least one functional group or moietywhich binds to the water-soluble polymer non-covalently (e.g., asdescribed herein in any one of the respective embodiments). In someembodiments, the water-soluble polymer is a polysaccharide (e.g., asdescribed herein in any one of the respective embodiments), and thelinker comprises at least one polysaccharide-binding polypeptide capableof selectively binding to the polysaccharide (e.g., as described hereinin any one of the respective embodiments). In some embodiments, thewater-soluble polymer is hyaluronic acid (e.g., as described herein inany one of the respective embodiments), and the linker comprises atleast one hyaluronic acid-binding polypeptide capable of selectivelybinding to the hyaluronic acid (e.g., as described herein in any one ofthe respective embodiments).

In some embodiments of any one of the embodiments described herein,attachment of the water-soluble polymer to the surface comprisesattaching the linker to the surface covalently, thereby forming amodified surface to which the water-soluble polymer is attachable. Sucha modified surface comprises at least one functional group or moietycapable of binding to the water-soluble polymer non-covalently. In someembodiments, a method described herein comprises attaching the linker tothe surface prior to effecting attachment of the water-soluble polymerto the resulting modified surface.

In some embodiments of any one of the embodiments described herein,attachment of the water-soluble polymer to the surface comprisesattaching the linker to the water-soluble polymer non-covalently,thereby forming a modified water-soluble polymer which is attachable tothe surface. Such a modified water-soluble polymer comprises at leastone functional group or moiety capable of binding to the surfacecovalently. In some embodiments, a method described herein comprisesattaching the linker to the water-soluble polymer prior to effectingattachment of the resulting modified water-soluble polymer to thesurface.

As used herein, the phrase “polysaccharide-binding polypeptide”encompasses any polypeptide or oligopeptide (e.g., peptide chains of atleast 4 amino acid residues in length) capable of selectively binding(e.g., non-covalently) to a polysaccharide. A wide variety ofpolysaccharide-binding polypeptides and their binding specificities willbe known to the skilled person, and include short peptide sequences(e.g., from 4 to 50, optionally 4 to 20 amino acid residues in length),and longer polypeptides such as proteins or fragments (e.g.,carbohydrate-binding modules and/or domains) thereof. In addition, thephrase “polysaccharide-binding polypeptide” encompasses antibodiescapable of specifically binding to a polysaccharide. Such antibodieswill be available to the skilled person and/or the skilled person willknow how to prepare such antibodies, using immunological techniquesknown in the art.

Examples of polysaccharide-binding polypeptides which may be used insome of any one of the embodiments of the invention include, withoutlimitation, carbohydrate-binding modules (CBMs); and hyaluronicacid-binding peptides, polypeptides and/or modules (e.g., having asequence as described in any of International Patent Applicationpublication WO 2013/110056; International Patent Application publicationWO 2014/071132; Barta et al. [Biochem J 1993, 292:947-949], Kohda et al.[Cell 1996, 86:767-775], Brisset & Perkins [FEBS Lett 1996,388:211-216], Peach et al. [J Cell Biol 1993, 122:257-264] and Zaleskiet al. [Antimicrob Agents Chemother 2006, 50:3856-3860], the contents ofwhich are incorporated herein by reference in their entirety).

Examples of CBMs which may be used in some of any one of the embodimentsof the invention, include, without limitation, CBMs belonging to thefamilies CBM3, CBM4, CBM9, CBM10, CBM17 and/or CBM28 (which mayoptionally be used to bind cellulose, e.g., in a surface); CBMS, CBM12,CBM14, CBM18, CBM19 and/or CBM33 (which may optionally be used to bindchitin and/or other polysaccharides comprising N-acetylglucosamine,e.g., in some of the water-soluble polymers described herein); CBM15(which may optionally be used to bind hemicellulose, e.g., in awood-based surface); and/or CBM20, CBM21 and/or CBM48 (which mayoptionally be used to bind starch and/or glycogen).

It is expected that during the life of a patent maturing from thisapplication many relevant functional groups and moieties for bindingwill be developed and/or uncovered and the scope of the terms“functional group”, “moiety”, “linker” and “polysaccharide-bindingpolypeptide” and the like is intended to include all such newtechnologies a priori.

In some embodiments of any one of the embodiments described herein, thewater-soluble polymer is attached (e.g., covalently attached) to thesurface, as described herein in any one of the respective embodiments,prior to contact of the water-soluble polymer and/or surface with theliposomes, thereby limiting reaction of the liposomes with reactivefunctional groups of the (modified or non-modified) water-solublepolymer and/or surface. In some embodiments, the water-soluble polymerand surface are essentially devoid of functional groups capable ofcovalently binding to the liposomes, when the liposomes are contactedwith the water-soluble polymer and surface.

In some embodiments of any one of the embodiments described herein, thewater-soluble polymer is attached (e.g., covalently attached) to thesurface, as described herein in any one of the respective embodiments,concomitantly and/or subsequent to contact of the water-soluble polymerwith liposomes, for example, embodiments in which the surface (modifiedor non-modified, as described herein in any one of the respectiveembodiments) is contacted with a solution comprising a water-solublepolymer (modified or non-modified, as described herein in any one of therespective embodiments), liposomes, and an aqueous carrier (e.g., asdescribed herein in any one of the respective embodiments). In some suchembodiments, the water-soluble polymer and surface (and optionally alsothe liposomes) are selected such that the water-soluble polymer isattachable to the surface in the presence of liposomes, that is, theliposomes do not interfere with attachment (e.g., covalent attachment)of the water-soluble polymer to the surface.

In some embodiments of any one of the embodiments described herein, thewater-soluble polymer is attached (e.g., covalently attached) to thesurface, as described herein in any one of the respective embodiments,concomitantly and/or subsequent to contact of the water-soluble polymerwith liposomes, and a functional group on the water-soluble polymer forattaching the water-soluble polymer to the surface is selected so as notto be attachable to the liposome lipids. For example, the water-solublepolymer (e.g., modified water-soluble polymer) may optionally comprise afunctional group which forms a covalent bond with a nucleophilic groupdescribed herein, and the lipids are selected so as to not covalentlyreact with such a functional group.

In some embodiments of any one of the embodiments described herein, thewater-soluble polymer is attached (e.g., covalently attached) to thesurface, as described herein in any one of the respective embodiments,concomitantly and/or subsequent to contact of the surface withliposomes, and a functional group on the surface for attaching thewater-soluble polymer to the surface is selected so as not to beattachable to the liposome lipids. For example, the surface (e.g.,modified surface) may optionally comprise a functional group which formsa covalent bond with a nucleophilic group described herein, and thelipids are selected so as to not covalently react with such a functionalgroup.

Phosphatidylcholines are examples of lipids which do not have a reactivenucleophilic group as described herein, whereas the similarphosphatidylethanolamines comprise a primary amine group which may reactwith a number of functional groups as described herein.

Composition-of-Matter:

According to another aspect of embodiments of the invention, there isprovided a composition-of-matter comprising a substrate coated, on atleast a portion of a surface thereof, by at least one water-solublepolymer. The water-soluble polymer(s) on the surface is coated by anamphiphilic lipid comprising at least one hydrophilic group.

According to another aspect of embodiments of the invention, there isprovided an article of manufacture comprising a composition-of-matteraccording to any one of the embodiments described herein.

According to another aspect of embodiments of the invention, there isprovided an article of manufacture comprising a composition-of-matter,the composition-of-matter comprising a substrate coated, on at least aportion of a surface thereof, by at least one water-soluble polymer, thearticle of manufacture being identified for use for efficientlyattaching thereto an amphiphilic lipid so as to reduce a frictioncoefficient of said substrate (e.g., according to any of the respectiveembodiments described herein relating to attaching a lipid and/orreducing a friction coefficient). Herein, the term“composition-of-matter” refers to any composition comprising a pluralityof substances (e.g., substrate, water-soluble polymer(s), amphiphiliclipid) in a form which does not exist in nature, and which does notinclude a portion of a human being. The form which does not exist innature may optionally comprise natural substances in a combination whichdoes not exist in nature, and/or may optionally comprise one or moresubstances which do not occur in nature. It is to be understood thatthis definition is not necessarily identical with a standard legaldefinition of the term.

Herein, the term “article of manufacture” refers to any article producedfrom materials in a manner which results in new forms, qualities,properties or combinations of the materials. It is to be understood thatthis definition is not necessarily identical with a standard legaldefinition of the term. The article of manufacture described herein mayoptionally consist essentially of the composition-of-matter, oralternatively, may comprise additional materials and/or parts.

In some embodiments of any one of the embodiments described herein, theamphiphilic lipid comprises at least one charged group (e.g., one ormore negatively charged groups and/or one or more positively chargedgroups).

In some embodiments, the amphiphilic lipid is zwitterionic, comprisingan equal number of negatively charged and positively charged groups(e.g., one of each).

A composition-of-matter according to embodiments of any of the aspectsdescribed in this section may include an amphiphilic lipid according toany one of the embodiments described herein with respect to liposomesand/or lipids, and water-soluble polymer(s) according to any one of theembodiments described herein with respect to water-soluble polymer(s).In addition, the water-soluble polymer(s) may be attached to thesubstrate according to any one of the embodiments described herein withrespect to attachment of the water-soluble polymer(s) to a surface. Insome embodiments, the water-soluble polymer(s) is attached to thesubstrate via a linker, as described herein in any one of the respectiveembodiments.

Thus, at least a portion of the composition-of-matter exhibits a layeredstructure, with the layers being in the order substrate-water-solublepolymer-amphiphilic lipid.

It is to be appreciated that the water-soluble polymer(s) may be in aform a very thin layer, and does not need to be in a bulk form (e.g.,gel).

Indeed, without being bound by any particular theory, it is believedthat a very thin layer may in many embodiments be more robust than abulk form such as a gel, for example, with respect to high appliedpressures.

In some embodiments of any one of the embodiments described herein, anaverage thickness of a layer of water-soluble polymer(s) on the surfaceis no more than 1 μm. In some embodiments, the average thickness is nomore than 300 nm. In some embodiments, the average thickness is no morethan 300 nm. In some embodiments, the thickness is no more than 100 nm.In some embodiments, the average thickness is no more than 30 nm. Insome embodiments, the average thickness is no more than 10 nm. In someembodiments, the average thickness is no more than 3 nm. In exemplaryembodiments, the average thickness is no more than 1.5 nm, the thicknessbeing in a range of about 0.3-1.5 nm.

At least a portion of the molecules of the amphiphilic lipid areoriented such that hydrophilic groups thereof (e.g., charged groups)face outwards at a surface of the composition-of-matter. In someembodiments of any one of the embodiments described herein, at least 50%of the molecules are oriented such that hydrophilic groups (e.g.,charged groups) face outwards. In some embodiments, at least 70% of themolecules are oriented such that hydrophilic groups (e.g., chargedgroups) face outwards. In some embodiments, at least 90% of themolecules are oriented such that hydrophilic groups (e.g., chargedgroups) face outwards. In some embodiments, at least 95% of themolecules are oriented such that hydrophilic groups (e.g., chargedgroups) face outwards. In some embodiments, at least 98% of themolecules are oriented such that hydrophilic groups (e.g., chargedgroups) face outwards. In some embodiments, at least 99% of themolecules are oriented such that hydrophilic groups (e.g., chargedgroups) face outwards.

As used herein, the phrase “face outwards at a surface” refers to agroup in a molecule (e.g., a lipid) which is closer to the surface ofthe composition-of-matter than the center of gravity of the molecule isto the surface of the composition-of-matter, and farther from thesubstrate than the center of gravity of the molecule is from thesubstrate.

As discussed herein, and without being bound by any particular theory,it is believed that outwards facing hydrophilic groups (e.g., chargedgroups) according to embodiments of the invention effect highlyeffective lubrication due, at least in part, to hydration lubricationassociated with hydrated hydrophilic groups, especially hydrated chargedgroups.

In some embodiments of any one of the embodiments described herein, atleast a portion of the amphiphilic lipid is in a form of a bilayer, thebilayer having a lipophilic region (e.g., a layer consisting primarilyof lipophilic moieties of the lipids) between two hydrophilic regions(e.g., hydrophilic layers) which comprise hydrophilic moieties (e.g.,charged groups) of the lipids, that is, the lipophilic region issandwiched between two hydrophilic regions.

The amphiphilic lipids in a bilayer are optionally oriented such thathydrophilic groups (e.g., charged groups) of lipids on the external sideof the bilayer face outwards (at the surface of thecomposition-of-matter), and hydrophilic groups (e.g., charged groups) oflipids on the internal side of the bilayer face inwards, that is,towards the water-soluble polymer(s) and substrate, and the lipophilicmoieties (e.g., fatty acyl groups) of the lipids on both sides of thebilayer (the internal and external sides) meet in the middle of thebilayer (e.g., thereby forming the lipophilic region of the bilayer). Insome embodiments of any one of the embodiments described herein, abilayer is bound to the water-soluble polymer(s) by electrostaticattraction. The electrostatic attraction may comprise attraction betweena pair of charged groups (e.g., an ionic bond), between an ionic groupand a dipole and/or between two dipoles. A dipole involved in theelectrostatic attraction may comprise, for example, a dipole of anon-ionic atom or group (e.g., hydroxy, amine) in a water-solublepolymer (e.g., non-ionic polymer) and/or a dipole of a zwitterion (e.g.,a negatively charged group near a positively charged group, such as inphosphocholine).

In some embodiments of any one of the embodiments described herein, atleast a portion of the amphiphilic lipid is in a form of a monolayer,which may optionally be interspersed among a bilayer. The monolayer hasa lipophilic surface which comprises lipophilic moieties of the lipidsand a hydrophilic surface which comprises hydrophilic moieties (e.g.,charged groups) of the lipids.

The amphiphilic lipids in a monolayer are optionally oriented such thatthe hydrophilic surface of the monolayer faces outwards (at the surfaceof the composition-of-matter), and the lipophilic surface of themonolayer faces inwards, that is, towards the water-soluble polymer(s)and substrate. In some embodiments, a monolayer is bound to thewater-soluble polymer(s) and/or substrate by a hydrophobic interaction.In some embodiments, a distribution of the monolayer in the coatedsubstrate is associated with lipophilic regions in the water-solublepolymer(s) and/or gaps in the water-soluble polymer(s) which expose aregion (e.g., lipophilic region) of the substrate, with lipids in otherregions in the coated substrate being in a form other than a monolayer(e.g., a lipid bilayer, as described herein in any one of the respectiveembodiments).

In any of the embodiments described herein, the substrate may compriseany type of material or combination of different types of material,including inorganic material and/or organic material, in crystalline,amorphous and/or gel (e.g., hydrogel) forms, for example, metal,mineral, ceramic, glass, polymer (e.g., synthetic polymer, biopolymer),plant and/or animal biomass, and combinations thereof.

In some embodiments, the substrate comprises a physiological surface(e.g., a physiological tissue) and/or a surface in contact with and/orintended to come into contact with a physiological surface (e.g., asdescribed herein in any one of the respective embodiments).

Physiological Environment:

In some embodiments of any one of the aspects described herein, thesubstrate and/or surface described herein is a physiological surface,and/or a surface in contact with and/or intended to come into contactwith a physiological surface.

Any one of the embodiments described herein relating to a method ofreducing a friction coefficient of a surface and/or a surface coatedwith at least one water-soluble polymer and an amphiphilic lipid mayoptionally be further limited according to any one of the embodiments inthis section.

Herein, the phrase “physiological surface” refers to a surface of a partof a body.

A surface in contact with and/or intended to come into contact with aphysiological surface may be, for example, an implant, and/or a suture.

Without being bound by any particular theory, it is believed that themethod described herein is particularly suitable for application tophysiological surfaces or surfaces which come into contact them, becausethe liposomes and water-soluble polymer(s) may readily be selected so asto be biocompatible, optionally even substances naturally occurring inthe body, and because hydration lubrication mechanism (e.g., asdescribed herein in any one of the respective embodiments) is fullycompatible with aqueous environments such as physiological environments,as opposed, for example, to lubrication via non-aqueous liquidlubricants (e.g., oils).

In some embodiments of any one of the embodiments described herein, thesurface is physiological surface of a joint (e.g., an articular surface)and/or a surface in contact with and/or intended to come into contactwith a physiological surface of a joint (e.g., a joint implant). In someembodiments, the joint is a synovial joint.

In some embodiments of any one of the embodiments described herein, thephysiological surface comprises cartilage. In some embodiments, thecartilage is articular cartilage.

In some embodiments according to any of the embodiments described hereinrelating to reducing a friction coefficient of a surface in a joint(e.g., an articular surface of a synovial joint), the liposomes areselected such that the lipids on the surface are in a solid phase in thejoint (e.g., under physiological conditions).

Without being bound by any particular theory, it is believed that thesolid phase is more robust the liquid phase, particularly at therelatively high pressures to which articular surfaces are commonlysubjected.

In some embodiments of any one of the embodiments described herein, theliposomes are characterized by a phase transition melting point (Tm)above 37° C. In some embodiments, the Tm is above 38° C. In someembodiments, the Tm is above 39° C. In some embodiments, the Tm is above40° C. In some embodiments, the Tm is above 42° C. In some embodiments,the Tm is above 45° C. In some embodiments, the Tm is above 50° C. Insome embodiments, the Tm is above 55° C.

In some embodiments of any one of the embodiments described herein,attaching water-soluble polymer(s) to a physiological surface (e.g., anarticular surface of a synovial joint) is effected by parenteraladministration of an aqueous solution of the water-soluble polymer(s).The aqueous solution optionally comprises a physiologically acceptablecarrier.

In some embodiments of any one of the embodiments described herein,contacting a water-soluble polymer with liposomes is effected in thevicinity of a physiological surface (e.g., an articular surface of asynovial joint) by parenteral administration of an aqueous solution ofthe liposomes. The aqueous solution optionally comprises aphysiologically acceptable carrier.

In some embodiments of any one of the embodiments described herein, asolution comprising the water-soluble polymer(s) is administered (e.g.,as described herein in any one of the respective embodiments), andsubsequently, a solution comprising the liposomes is administered (e.g.,as described herein in any one of the respective embodiments).

In some embodiments of any one of the embodiments described herein, thewater-soluble polymer(s) and liposomes are administered concomitantly.

In some embodiments of any one of the embodiments described herein, themethod comprises contacting a physiological surface (e.g., an articularsurface of a synovial joint) with a solution comprising water-solublepolymer(s), liposomes and an aqueous carrier (e.g., as described hereinin any one of the respective embodiments) via parenteral administration.The aqueous carrier is optionally a physiologically acceptable carrier.

In some embodiments of any one of the embodiments described herein, themethod comprises modifying a physiological surface, for example, anarticular surface of a synovial joint (e.g., as described herein in anyone of the respective embodiments), thereby resulting in a modifiedphysiological surface to which the water-soluble polymer is attachable(e.g., as described herein in any one of the respective embodiments). Insome such embodiments, the modifying is effected with a solutioncomprising a reagent (e.g., a linker described herein) and an aqueouscarrier (e.g., as described herein in any one of the respectiveembodiments) via parenteral administration. The aqueous carrier isoptionally a physiologically acceptable carrier.

In some embodiments of any one of the embodiments described herein, themethod comprises modifying a water-soluble polymer (e.g., as describedherein in any one of the respective embodiments), thereby resulting in amodified water-soluble polymer attachable to a physiological surface,for example, an articular surface of a synovial joint. In some suchembodiments, the modifying is effected with a solution comprising themodified water-soluble polymer, liposomes and an aqueous carrier (e.g.,as described herein in any one of the respective embodiments) viaparenteral administration. In such embodiments, the modification whichenhances attachability does not require any additional treatment step,as the modification may performed (on the water-soluble polymer) exvivo.

In some embodiments of any one of the embodiments described herein,parenteral administration of any of the solutions described hereincomprises injection of a solution described herein solution into thevicinity of the surface. In some embodiments, the surface is anarticular surface of a synovial joint and the solution is injected intothe synovial cavity.

In some embodiments of any one of the embodiments described herein, thewater-soluble polymer (e.g., hyaluronic acid) is attachable to thephysiological surface by covalent binding and/or by selectivenon-covalent binding to collagen (e.g., as described herein in any oneof the respective embodiments).

In some embodiments of any one of the embodiments described herein, thewater-soluble polymer is a modified water-soluble polymer (e.g., asdescribed herein in any one of the respective embodiments), in which awater-soluble polymer (e.g., hyaluronic acid) is modified so as tofurther comprise at least one functional group (e.g., a dihydroxyphenylgroup described herein) for attaching the polymer to collagen.

In some embodiments of any one of the embodiments described herein, themodified or non-modified water-soluble polymer (e.g., hyaluronic acid)comprises at least one functional group which forms a covalent bond withamine groups (e.g., as described herein in any one of the respectiveembodiments) which are present on the physiological surface (e.g., aminegroups of polypeptides, such as lysine residues, on the physiologicalsurface). In some embodiments, the water-soluble polymer (e.g., modifiedwater-soluble polymer) comprises at least one dihydroxyphenyl group(e.g., as described herein in any one of the respective embodiments). Insome embodiments, the water-soluble polymer (e.g., modifiedwater-soluble polymer) comprises at least one nucleophilic leaving group(e.g., as described herein in any one of the respective embodiments). Insome embodiments, the water-soluble polymer (e.g., modifiedwater-soluble polymer) comprises at least one N-hydroxysuccinimideleaving group.

In some embodiments of any one of the embodiments described herein,attachment of the water-soluble polymer to the physiological surface iseffected via a linker (e.g., a linker as described herein in any one ofthe respective embodiments). In some embodiments, the linker is adaptedfor attaching a water-soluble polymer which is a polysaccharide tocollagen. In some embodiments, the linker is adapted for attachinghyaluronic acid to collagen. In some embodiments, the collagen is typeII collagen, also referred to as collagen II (a type of collagen whichis abundant in articular cartilage).

Examples of functional groups or moieties which may optionally beincluded in a linker in order to effect attachment to articularcartilage and/or collagen include, without limitation, functional groupswhich form covalent bonds with amine groups as described herein (e.g.,amine groups are abundant in collagen and cartilage); and moietiescapable of selectively binding collagen (e.g., collagen II)non-covalently, such as collagen-binding polypeptides (e.g., collagenII-binding polypeptides).

As used herein, the phrase “collagen-binding polypeptide” encompassesany polypeptide or oligopeptide (e.g., peptide chains of at least 4amino acid residues in length) capable of selectively binding (e.g.,non-covalently) to a collagen (e.g., one type of collagen, some types ofcollagen, all types of collagen), including glycosylated polypeptidesand oligopeptides such as peptidoglycans and proteoglycans. A widevariety of collagen-binding polypeptides and their binding specificitieswill be known to the skilled person, and include short peptide sequences(e.g., from 4 to 50, optionally 4 to 20 amino acid residues in length),and longer polypeptides such as proteins or fragments (e.g.,collagen-binding domains) thereof. In addition, the phrase“collagen-binding polypeptide” encompasses antibodies capable ofspecifically binding to a collagen. Such antibodies will be available tothe skilled person and/or the skilled person will know how to preparesuch antibodies, using immunological techniques known in the art.

Examples of collagen-binding polypeptides which may be used inembodiments of the invention include, without limitation,collagen-binding proteins (e.g., decorin), fragments thereof and/orother polypeptides as described in U.S. Pat. No. 8,440,618, Abd-Elgaliel& Tung [Biopolymers 2013, 100:167-173], Paderi et al. [Tissue Eng Part A2009, 15:2991-2999], Rothenfluh et al. [Nat Mater 2008, 7:248-254] andHelms et al. [J Am Chem Soc 2009, 131:11683-11685] (the contents of eachof which are incorporated herein by reference in their entirety).

It is expected that during the life of a patent maturing from thisapplication many relevant collagen-binding polypeptides will bedeveloped and/or uncovered and the scope of the term “collagen-bindingpolypeptide” is intended to include all such new technologies a priori.

In some embodiments of any one of the embodiments described herein, thelinker comprises at least one functional group or moiety which binds tothe physiological surface (e.g., articular cartilage) non-covalently(e.g., as described herein in any one of the respective embodiments),and at least one functional group or moiety which binds to thewater-soluble polymer non-covalently (e.g., as described herein in anyone of the respective embodiments). In some embodiments, the linkercomprises a moiety capable of selectively binding collagen (e.g.,collagen II) non-covalently, e.g., as described herein in any one of therespective embodiments. In some embodiments, the linker comprises acollagen-binding polypeptide (e.g., a collagen II-binding polypeptide),e.g., as described herein in any one of the respective embodiments.

In some embodiments, attachment of the water-soluble polymer to thephysiological surface (e.g., articular cartilage) comprises attachingthe linker to the physiological surface non-covalently, thereby forminga modified physiological surface to which the water-soluble polymer isattachable. Such a modified physiological surface comprises at least onefunctional group or moiety capable of binding to the water-solublepolymer non-covalently. In some embodiments, a method described hereincomprises attaching the linker to the physiological surface (e.g.,articular cartilage) prior to effecting attachment of the water-solublepolymer to the resulting modified physiological surface.

In some embodiments, attachment of the water-soluble polymer to thephysiological surface (e.g., articular cartilage) comprises attachingthe linker to the water-soluble polymer non-covalently, thereby forminga modified water-soluble polymer which is attachable to thephysiological surface. Such a modified water-soluble polymer comprisesat least one functional group or moiety capable of binding to thephysiological surface non-covalently. In some embodiments, a methoddescribed herein comprises attaching the linker to the water-solublepolymer prior to effecting attachment of the resulting modifiedwater-soluble polymer to the physiological surface (e.g., articularcartilage).

In some embodiments of any one of the embodiments described herein, thelinker comprises at least one functional group or moiety which binds tothe physiological surface (e.g., articular cartilage) covalently (e.g.,as described herein in any one of the respective embodiments), and atleast one functional group or moiety which binds to the water-solublepolymer covalently (e.g., as described herein in any one of therespective embodiments). In some embodiments, the linker comprises afunctional group which forms a covalent bond with an amine group (e.g.,as described herein in any one of the respective embodiments). In someembodiments, the linker comprises at least one dihydroxyphenyl group. Insome embodiments, the linker comprises at least one nucleophilic leavinggroup. In some embodiments, the linker comprises at least oneN-hydroxysuccinimide leaving group.

In some embodiments, attachment of the water-soluble polymer to thephysiological surface (e.g., articular cartilage) comprises attachingthe linker to the physiological surface covalently, thereby forming amodified physiological surface to which the water-soluble polymer isattachable. Such a modified physiological surface comprises at least onefunctional group or moiety capable of binding to the water-solublepolymer covalently. In some embodiments, a method described hereincomprises attaching the linker to the physiological surface (e.g.,articular cartilage) prior to effecting attachment of the water-solublepolymer to the resulting modified physiological surface.

In some embodiments, attachment of the water-soluble polymer to thephysiological surface (e.g., articular cartilage) comprises attachingthe linker to the water-soluble polymer covalently, thereby forming amodified water-soluble polymer which is attachable to the physiologicalsurface. Such a modified water-soluble polymer comprises at least onefunctional group or moiety capable of binding to the physiologicalsurface covalently. In some embodiments, a method described hereincomprises attaching the linker to the water-soluble polymer prior toeffecting attachment of the resulting modified water-soluble polymer tothe physiological surface (e.g., articular cartilage).

In some embodiments of any one of the embodiments described herein, thelinker comprises at least one functional group or moiety which binds tothe physiological surface (e.g., articular cartilage) non-covalently(e.g., as described herein in any one of the respective embodiments),and at least one functional group or moiety which binds to thewater-soluble polymer covalently (e.g., as described herein in any oneof the respective embodiments). In some embodiments, the linkercomprises a moiety capable of selectively binding collagen (e.g.,collagen II) non-covalently, e.g., as described herein in any one of therespective embodiments. In some embodiments, the linker comprises acollagen-binding polypeptide (e.g., a collagen II-binding polypeptide),e.g., as described herein in any one of the respective embodiments.

In some embodiments, attachment of the water-soluble polymer to thephysiological surface (e.g., articular cartilage) comprises attachingthe linker to the physiological surface non-covalently, thereby forminga modified physiological surface to which the water-soluble polymer isattachable. Such a modified physiological surface comprises at least onefunctional group or moiety capable of binding to the water-solublepolymer covalently. In some embodiments, a method described hereincomprises attaching the linker to the physiological surface (e.g.,articular cartilage) prior to effecting attachment of the water-solublepolymer to the resulting modified physiological surface.

In some embodiments, attachment of the water-soluble polymer to thephysiological surface (e.g., articular cartilage) comprises attachingthe linker to the water-soluble polymer covalently, thereby forming amodified water-soluble polymer which is attachable to the physiologicalsurface. Such a modified water-soluble polymer comprises at least onefunctional group or moiety capable of binding to the physiologicalsurface non-covalently. In some embodiments, a method described hereincomprises attaching the linker to the water-soluble polymer prior toeffecting attachment of the resulting modified water-soluble polymer tothe physiological surface (e.g., articular cartilage).

In some embodiments of any one of the embodiments described herein, thelinker comprises at least one functional group or moiety which binds tothe physiological surface (e.g., articular cartilage) covalently (e.g.,as described herein in any one of the respective embodiments), and atleast one functional group or moiety which binds to the water-solublepolymer non-covalently (e.g., as described herein in any one of therespective embodiments). In some embodiments, the linker comprises afunctional group which forms a covalent bond with an amine group (e.g.,as described herein in any one of the respective embodiments). In someembodiments, the linker comprises at least one dihydroxyphenyl group. Insome embodiments, the linker comprises at least one nucleophilic leavinggroup. In some embodiments, the linker comprises at least oneN-hydroxysuccinimide leaving group.

In some embodiments, attachment of the water-soluble polymer to thephysiological surface (e.g., articular cartilage) comprises attachingthe linker to the physiological surface covalently, thereby forming amodified physiological surface to which the water-soluble polymer isattachable. Such a modified physiological surface comprises at least onefunctional group or moiety capable of binding to the water-solublepolymer non-covalently. In some embodiments, a method described hereincomprises attaching the linker to the physiological surface (e.g.,articular cartilage) prior to effecting attachment of the water-solublepolymer to the resulting modified physiological surface.

In some embodiments, attachment of the water-soluble polymer to thephysiological surface (e.g., articular cartilage) comprises attachingthe linker to the water-soluble polymer non-covalently, thereby forminga modified water-soluble polymer which is attachable to thephysiological surface. Such a modified water-soluble polymer comprisesat least one functional group or moiety capable of binding to thephysiological surface covalently. In some embodiments, a methoddescribed herein comprises attaching the linker to the water-solublepolymer prior to effecting attachment of the resulting modifiedwater-soluble polymer to the physiological surface (e.g., articularcartilage).

In some embodiments of any one of the embodiments described herein, thewater-soluble polymer is a polysaccharide (e.g., as described herein inany one of the respective embodiments), and the linker comprises atleast one polysaccharide-binding polypeptide capable of selectivelybinding to the polysaccharide (e.g., as described herein in any one ofthe respective embodiments), thereby effecting attachment to thephysiological surface (e.g., articular cartilage). In some embodiments,the water-soluble polymer is hyaluronic acid (e.g., as described hereinin any one of the respective embodiments), and the linker comprises atleast one hyaluronic acid-binding polypeptide capable of selectivelybinding to the hyaluronic acid (e.g., as described herein in any one ofthe respective embodiments).

In some embodiments according to any of the embodiments described hereinrelating to reducing a friction coefficient of a surface in a joint, themethod and/or solution described herein for reducing a frictioncoefficient is for use in the treatment of a synovial joint disorderassociated with an increased friction coefficient of an articularsurface in the synovial joint.

According to another aspect of embodiments of the invention, there isprovided a use of a solution for reducing a friction coefficient of asurface, as described herein in any one of the respective embodiments,in the manufacture of a medicament for treating a synovial jointdisorder associated with an increased friction coefficient of anarticular surface in the synovial joint.

Examples of synovial joint disorders associated with an increasedfriction coefficient of an articular surface, and treatable according toembodiments of various aspects of the invention, include, withoutlimitation, arthritis, traumatic joint injury, locked joint (also knownin the art as joint locking), and joint injury associated with surgery.

In some embodiments, the arthritis is selected from the group consistingof osteoarthritis, rheumatoid arthritis and psoriatic arthritis.

In some embodiments, the locked joint is associated with osteochondritisdissecans and/or synovial osteochondromatosis.

The joint injury associated with surgery described herein may optionallybe associated with surgery which directly inflicts damage on anarticular surface (e.g., by incision), and/or surgery which damages anarticular surface only indirectly. For example, surgery which repairs orotherwise affects tissue in the vicinity of the joint (e.g., ligamentsand/or menisci) may be associated with joint injury due to alteredmechanics in the joint.

The traumatic joint injury described herein may optionally be injurycaused directly by trauma (e.g., inflicted at the time of the trauma)and/or injury caused by previous trauma (e.g., a post-traumatic injurywhich develops sometime after the trauma).

The water-soluble polymer(s) and/or liposomes may optionally beadministered as part of a solution that comprises a physiologicallyacceptable carrier, for example an aqueous carrier which is aphysiologically acceptable carrier.

Herein, the term “physiologically acceptable carrier” refers to acarrier or a diluent that does not cause significant irritation to asubject upon administration in the intended manner, and does notabrogate the activity and properties of the water-soluble polymer(s)and/or liposomes in the solution (e.g., their ability to reduce afriction coefficient of a surface, as described herein in any one of therespective embodiments). Examples, without limitations, of carriers are:propylene glycol, saline, emulsions and mixtures of organic solventswith water, as well as solid (e.g., powdered) and gaseous carriers.

Techniques for formulation and administration of compounds may be foundin “Remington's Pharmaceutical Sciences” Mack Publishing Co., Easton,Pa., latest edition, which is incorporated herein by reference.

Solutions according to any one of the embodiments of the presentinvention may be manufactured by processes well known in the art, e.g.,by means of conventional mixing or dissolving processes.

Solutions for use in accordance with the present invention thus may beformulated in conventional manner using one or more physiologicallyacceptable carriers, which facilitate processing of the water-solublepolymer(s) and/or liposomes into preparations which can be usedpharmaceutically. Proper formulation is dependent upon the route ofadministration chosen.

For injection, the water-soluble polymer(s) and/or liposomes describedherein may be formulated in aqueous solutions, preferably inphysiologically compatible buffers such as Hank's solution, Ringer'ssolution, histidine buffer, or physiological saline buffer with orwithout organic solvents such as propylene glycol, polyethylene glycol.

The water-soluble polymer(s) and/or liposomes described herein may beformulated for parenteral administration, e.g., by bolus injection orcontinuous infusion. Formulations for injection may be presented in unitdosage form, e.g., in ampoules or in multidose containers withoptionally, an added preservative. The compositions may be suspensions,solutions or emulsions in oily or aqueous vehicles, and may containformulatory agents such as suspending, stabilizing and/or dispersingagents.

The water-soluble polymer(s) and/or liposomes described herein may beformulated as an aqueous solution per se. Additionally, the solution maybe in the form of a suspension and/or emulsions (e.g., the aqueous phaseof a suspension or water-in-oil, oil-in-water or water-in-oil-in-oilemulsion), for example, in order to increase the viscosity of theformulation. Aqueous injection suspensions may contain substances, whichincrease the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol or dextran. Optionally, the suspension may alsocontain suitable stabilizers or agents, which increase the solubility ofthe water-soluble polymer(s) and/or liposomes described herein, forexample, to allow for the preparation of highly concentrated solutions.

In some embodiments, the water-soluble polymer(s) and/or liposomesdescribed herein may be in powder form for constitution with a suitablevehicle, e.g., sterile, pyrogen-free water, before use.

The solutions may be formulated wherein the active ingredient(s)(water-soluble polymer(s) and/or liposomes) are contained in an amounteffective to achieve the intended purpose, for example, an amounteffective to prevent, alleviate or ameliorate symptoms of a disorder inthe subject being treated.

The dosage may vary depending upon the dosage form employed, the routeof administration utilized, and the location of administration (e.g.,the volume and/or surface of the region contacted with the water-solublepolymer(s) and/or liposomes).

The amount of a composition to be administered will, of course, bedependent on the subject being treated, the severity of the affliction,the manner of administration, the judgment of the prescribing physician,etc.

Solutions according to embodiments of the present invention may, ifdesired, be presented in a pack or dispenser device, such as an FDA (theU.S. Food and Drug Administration) approved kit, which may contain oneor more unit dosage forms containing the active ingredient(s) (e.g.,water-soluble polymer(s) and/or liposomes described herein). The packmay, for example, comprise metal or plastic foil, such as, but notlimited to a blister pack. The pack or dispenser device may beaccompanied by instructions for administration. The pack or dispensermay also be accompanied by a notice associated with the container in aform prescribed by a governmental agency regulating the manufacture, useor sale of pharmaceuticals, which notice is reflective of approval bythe agency of the form of the compositions for human or veterinaryadministration. Such notice, for example, may be of labeling approved bythe U.S. Food and Drug Administration for prescription drugs or of anapproved product insert. Solutions comprising water-soluble polymer(s)and/or liposomes, as described herein in any one of the respectiveembodiments, formulated in a physiologically acceptable carrier may alsobe prepared, placed in an appropriate container, and labeled fortreatment of an indicated condition or diagnosis, as is detailed herein.

Additional Definitions

Herein, the term “alkyl” describes a saturated aliphatic hydrocarbonincluding straight chain and branched chain groups. Preferably, thealkyl group has 1 to 20 carbon atoms. Whenever a numerical range; e.g.,“1 to 20”, is stated herein, it implies that the group, in this case thealkyl group, may contain 1 carbon atom, 2 carbon atoms, 3 carbon atoms,etc., up to and including 20 carbon atoms. More preferably, the alkyl isa medium size alkyl having 1 to 10 carbon atoms. Most preferably, unlessotherwise indicated, the alkyl is a lower alkyl having 1 to 4 carbonatoms. The alkyl group may be substituted or non-substituted.Substituted alkyl may have one or more substituents, whereby eachsubstituent group can independently be, for example, cycloalkyl,alkenyl, alkynyl, aryl, heteroaryl, heteroalicyclic, amine, halide,sulfonate, sulfoxide, phosphonate, hydroxy, alkoxy, aryloxy,thiohydroxy, thioalkoxy, thioaryloxy, cyano, nitro, azo, sulfonamide,carboxy, thiocarbamate, urea, thiourea, carbamate, amide, and hydrazine.

Herein, the term “alkenyl” describes an unsaturated aliphatichydrocarbon comprise at least one carbon-carbon double bond, includingstraight chain and branched chain groups. Preferably, the alkenyl grouphas 2 to 20 carbon atoms. More preferably, the alkenyl is a medium sizealkenyl having 2 to 10 carbon atoms. Most preferably, unless otherwiseindicated, the alkenyl is a lower alkenyl having 2 to 4 carbon atoms.The alkenyl group may be substituted or non-substituted. Substitutedalkenyl may have one or more substituents, whereby each substituentgroup can independently be, for example, cycloalkyl, alkynyl, aryl,heteroaryl, heteroalicyclic, amine, halide, sulfonate, sulfoxide,phosphonate, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy,thioaryloxy, cyano, nitro, azo, sulfonamide, carboxy, thiocarbamate,urea, thiourea, carbamate, amide, and hydrazine.

Herein, the term “alkynyl” describes an unsaturated aliphatichydrocarbon comprise at least one carbon-carbon triple bond, includingstraight chain and branched chain groups. Preferably, the alkynyl grouphas 2 to 20 carbon atoms. More preferably, the alkynyl is a medium sizealkynyl having 2 to 10 carbon atoms. Most preferably, unless otherwiseindicated, the alkynyl is a lower alkynyl having 2 to 4 carbon atoms.The alkynyl group may be substituted or non-substituted. Substitutedalkynyl may have one or more substituents, whereby each substituentgroup can independently be, for example, cycloalkyl, alkenyl, aryl,heteroaryl, heteroalicyclic, amine, halide, sulfonate, sulfoxide,phosphonate, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy,thioaryloxy, cyano, nitro, azo, sulfonamide, carboxy, thiocarbamate,urea, thiourea, carbamate, amide, and hydrazine.

The alkyl, alkenyl and/or alkynyl group can be an end group, as thisphrase is defined herein, wherein it is attached to a single adjacentatom, or a linking group, as this phrase is defined herein, whichconnects two or more moieties.

Herein, the phrase “end group” describes a group (e.g., a substituent)that is attached to a single moiety in the compound via one atomthereof.

The phrase “linking group” describes a group (e.g., a substituent) thatis attached to two or more moieties in the compound.

The term “cycloalkyl” describes an all-carbon monocyclic or fused ring(i.e., rings which share an adjacent pair of carbon atoms) group whereone or more of the rings does not have a completely conjugatedpi-electron system. The cycloalkyl group may be substituted ornon-substituted. Substituted cycloalkyl may have one or moresubstituents, whereby each substituent group can independently be, forexample, alkyl, cycloalkyl, aryl, heteroaryl, heteroalicyclic, amine,halide, sulfonate, sulfoxide, phosphonate, hydroxy, alkoxy, aryloxy,thiohydroxy, thioalkoxy, thioaryloxy, cyano, nitro, azo, sulfonamide,carboxy, thiocarbamate, urea, thiourea, carbamate, amide, and hydrazine.The cycloalkyl group can be an end group, as this phrase is definedherein, wherein it is attached to a single adjacent atom, or a linkinggroup, as this phrase is defined herein, connecting two or moremoieties.

The term “aryl” describes an all-carbon monocyclic or fused-ringpolycyclic (i.e., rings which share adjacent pairs of carbon atoms)groups having a completely conjugated pi-electron system. The aryl groupmay be substituted or non-substituted. Substituted aryl may have one ormore substituents, whereby each substituent group can independently be,for example, alkyl, cycloalkyl, aryl, heteroaryl, heteroalicyclic,amine, halide, sulfonate, sulfoxide, phosphonate, hydroxy, alkoxy,aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, cyano, nitro, azo,sulfonamide, carboxy, thiocarbamate, urea, thiourea, carbamate, amide,and hydrazine. The aryl group can be an end group, as this term isdefined herein, wherein it is attached to a single adjacent atom, or alinking group, as this term is defined herein, connecting two or moremoieties. Phenyl and naphthyl are representative aryl end groups.

The term “heteroaryl” describes a monocyclic or fused ring (i.e., ringswhich share an adjacent pair of atoms) group having in the ring(s) oneor more atoms, such as, for example, nitrogen, oxygen and sulfur and, inaddition, having a completely conjugated pi-electron system. Examples,without limitation, of heteroaryl groups include pyrrole, furan,thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrimidine,quinoline, isoquinoline and purine. The heteroaryl group may besubstituted or non-substituted. Substituted heteroaryl may have one ormore substituents, whereby each substituent group can independently be,for example, alkyl, cycloalkyl, aryl, heteroaryl, heteroalicyclic,amine, halide, sulfonate, sulfoxide, phosphonate, hydroxy, alkoxy,aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, cyano, nitro, azo,sulfonamide, carboxy, thiocarbamate, urea, thiourea, carbamate, amide,and hydrazine. The heteroaryl group can be an end group, as this phraseis defined herein, where it is attached to a single adjacent atom, or alinking group, as this phrase is defined herein, connecting two or moremoieties. Representative examples are pyridine, pyrrole, oxazole,indole, purine and the like.

The term “heteroalicyclic” describes a monocyclic or fused ring grouphaving in the ring(s) one or more atoms such as nitrogen, oxygen andsulfur. The rings may also have one or more double bonds. However, therings do not have a completely conjugated pi-electron system. Theheteroalicyclic may be substituted or non-substituted. Substitutedheteroalicyclic may have one or more substituents, whereby eachsubstituent group can independently be, for example, alkyl, cycloalkyl,aryl, heteroaryl, heteroalicyclic, amine, halide, sulfonate, sulfoxide,phosphonate, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy,thioaryloxy, cyano, nitro, azo, sulfonamide, carboxy, thiocarbamate,urea, thiourea, carbamate, amide, and hydrazine. The heteroalicyclicgroup can be an end group, as this phrase is defined herein, where it isattached to a single adjacent atom, or a linking group, as this phraseis defined herein, connecting two or more moieties. Representativeexamples are piperidine, piperazine, tetrahydrofuran, tetrahydropyran,morpholine and the like.

As used herein, the terms “amine” and “amino” describe both a —NRxRygroup —NRx- group, wherein Rx and Ry are each independently hydrogen,alkyl, cycloalkyl, aryl, heteroaryl or heteroalicyclic, as these termsare defined herein. When Rx or Ry is heteroaryl or heteroalicyclic, theamine nitrogen atom is bound to a carbon atom of the heteroaryl orheteroalicyclic ring.

The amine group can therefore be a primary amine, where both Rx and Ryare hydrogen, a secondary amine, where Rx is hydrogen and Ry is alkyl,cycloalkyl, aryl, heteroaryl or heteroalicyclic, or a tertiary amine,where each of Rx and Ry is independently alkyl, cycloalkyl, aryl,heteroaryl or heteroalicyclic.

The terms “halide” and “halo” refer to fluorine, chlorine, bromine oriodine. The term “haloalkyl” describes an alkyl group as defined herein,further substituted by one or more halide(s).

The term “phosphonate” refers to an —P(═O)(ORx)-OR_(Y) end group, or toa —P(═O)(ORx)-O— linking group, where Rx and RY are as defined herein.

The term “sulfoxide” or “sulfinyl” describes a —S(═O)—Rx end group or—S(═O)-linking group, where Rx is as defined herein.

The terms “sulfonate” and “sulfonyl” describe a —S(═O)₂—Rx end group or—S(═O)₂— linking group, where Rx is as defined herein.

The term “sulfonamide”, as used herein, encompasses both S-sulfonamideand N-sulfonamide end groups, and a —S(═O)₂—NRx- linking group.

The term “S-sulfonamide” describes a —S(═O)₂—NRxRY end group, with Rxand RY as defined herein.

The term “N-sulfonamide” describes an RxS(═O)₂—NR_(Y)— end group, whereRx and RY are as defined herein.

The term “carbonyl” as used herein, describes a —C(═O)—Rx end group or—C(═O) linking group, with Rx as defined herein.

The term “acyl” as used herein, describes a —C(═O)—Rx end group, with Rxas defined herein.

The terms “hydroxy” and “hydroxyl” describe a —OH group.

The term “alkoxy” describes both an —O-alkyl and an —O-cycloalkyl endgroup or linking group, as defined herein.

The term “aryloxy” describes both an —O-aryl and an —O-heteroaryl endgroup or linking group, as defined herein.

The term “thiohydroxy” describes a —SH group.

The term “thioalkoxy” describes both a —S-alkyl end group or linkinggroup, and a —S-cycloalkyl end group or linking group, as definedherein.

The term “thioaryloxy” describes both a —S-aryl and a —S-heteroaryl endgroup or linking group, as defined herein.

The terms “cyano” and “nitrile” describe a —C≡N group.

The term “nitro” describes an —NO₂ group.

The term “azo” describes an —N═N—Rx end group or —N═N═ linking group,with Rx as defined herein.

The terms “carboxy” and “carboxyl”, as used herein, encompasses bothC-carboxy and O-carboxy end groups, and a —C(═O)—O— linking group.

The term “C-carboxy” describes a —C(═O)—ORx end group, where Rx is asdefined herein.

The term “O-carboxy” describes a —OC(═O)—Rx end group, where Rx is asdefined herein.

The term “urea” describes a —NRxC(═O)—NRyRw end group or —NRxC(═O)—NRy-linking group, where Rx and Ry are as defined herein and Rw is asdefined herein for Rx and Ry.

The term “thiourea” describes a —NRx-C(═S)—NRyRw end group or a—NRx-C(═S)—NRy- linking group, with Rx, Ry and Ry as defined herein.

The term “amide”, as used herein, encompasses both C-amide and N-amideend groups, and a —C(═O)—NRx- linking group.

The term “C-amide” describes a —C(═O)—NRxRy end group, where Rx and Ryare as defined herein.

The term “N-amide” describes a RxC(═O)—NRy- end group, where Rx and Ryare as defined herein.

The term “carbamyl” or “carbamate”, as used herein, encompassesN-carbamate and O-carbamate end groups, and a —OC(═O)—NRx- linkinggroup.

The term “N-carbamate” describes an RyOC(═O)—NRx- end group, with Rx andRy as defined herein.

The term “O-carbamate” describes an —OC(═O)—NRxRy end group, with Rx andRy as defined herein.

The term “thiocarbamyl” or “thiocarbamate”, as used herein, encompassesboth O-thiocarbamate, S-thiocarbamate and N-thiocarbamate end groups,and a —OC(═S)—NRx- or —SC(═O)—NRx- linking group.

The term “O-thiocarbamate” describes a —OC(═S)—NRxRy end group, with Rxand Ry as defined herein.

The term “S-thiocarbamate” describes a —SC(═O)—NRxRy end group, with Rxand Ry as defined herein.

The term “N-thiocarbamate” describes an RyOC(═S)NRx- or RySC(═O)NRx-endgroup, with Rx and Ry as defined herein.

The term “guanidine” describes a —RxNC(═N)—NRyRw end group or—RxNC(═N)—NRy- linking group, where Rx, Ry and Rw are as defined herein.

The term “hydrazine”, as used herein, describes a —NRx-NRyRw end groupor —NRx-NRy- linking group, with Rx, Ry, and Rw as defined herein.

As used herein the term “about” refers to ±10%, and optionally ±5%. Theterms “comprises”, “comprising”, “includes”, “including”, “having” andtheir conjugates mean “including but not limited to”.

The term “consisting of” means “including and limited to”.

The term “consisting essentially of” means that the composition, methodor structure may include additional ingredients, steps and/or parts, butonly if the additional ingredients, steps and/or parts do not materiallyalter the basic and novel characteristics of the claimed composition,method or structure.

As used herein, the singular form “a”, “an” and “the” include pluralreferences unless the context clearly dictates otherwise. For example,the term “a compound” or “at least one compound” may include a pluralityof compounds, including mixtures thereof.

Throughout this application, various embodiments of this invention maybe presented in a range format. It should be understood that thedescription in range format is merely for convenience and brevity andshould not be construed as an inflexible limitation on the scope of theinvention. Accordingly, the description of a range should be consideredto have specifically disclosed all the possible subranges as well asindividual numerical values within that range. For example, descriptionof a range such as from 1 to 6 should be considered to have specificallydisclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numberswithin that range, for example, 1, 2, 3, 4, 5, and 6. This appliesregardless of the breadth of the range.

Whenever a numerical range is indicated herein, it is meant to includeany cited numeral (fractional or integral) within the indicated range.The phrases “ranging/ranges between” a first indicate number and asecond indicate number and “ranging/ranges from” a first indicate number“to” a second indicate number are used herein interchangeably and aremeant to include the first and second indicated numbers and all thefractional and integral numerals therebetween.

As used herein the term “method” refers to manners, means, techniquesand procedures for accomplishing a given task including, but not limitedto, those manners, means, techniques and procedures either known to, orreadily developed from known manners, means, techniques and proceduresby practitioners of the chemical, pharmacological, biological,biochemical and medical arts.

As used herein, the term “treating” includes abrogating, substantiallyinhibiting, slowing or reversing the progression of a condition,substantially ameliorating clinical or aesthetical symptoms of acondition or substantially preventing the appearance of clinical oraesthetical symptoms of a condition.

It is appreciated that certain features of the invention, which are, forclarity, described in the context of separate embodiments, may also beprovided in combination in a single embodiment. Conversely, variousfeatures of the invention, which are, for brevity, described in thecontext of a single embodiment, may also be provided separately or inany suitable sub-combination or as suitable in any other describedembodiment of the invention. Certain features described in the contextof various embodiments are not to be considered essential features ofthose embodiments, unless the embodiment is inoperative without thoseelements.

Various embodiments and aspects of the present invention as delineatedhereinabove and as claimed in the claims section below find experimentalsupport in the following examples.

EXAMPLES

Reference is now made to the following examples, which together with theabove descriptions illustrate some embodiments of the invention in anon-limiting fashion.

Example 1 Lubrication Solutions

Materials and Methods

Materials:

Hyaluronic acid (sodium hyaluronate, 1 and 1.5 MDa) was obtained fromLifecore Biomedical.

Phosphate buffer saline (PBS) was obtained from Sigma-Aldrich.

Phosphatidylcholines (PC), including dimyristoylphospatidylcholine(1,2-dimyristoyl-sn-glycero-3-phosphocholine; DMPC) and hydrogenated soyPC (HSPC), were obtained from Lipoid GmbH.

Polyethylene glycol (PEG or PEO), 200 kDa molecular weight, was obtainedfrom Sigma-Aldrich.

Polyvinylpyrrolidone (PVP), 40 kDa molecular weight, was obtained fromSigma-Aldrich.

Etafilcon A (1-Day ACUVUE®) and Narafilcon A (1-Day TruEye®) contactlenses were obtained from Johnson & Johnson, immersed in saline solutionin a blister-pack. The composition, water content and modulus of thecontact lenses are as follows. Etafilcon A lenses contain2-hydroxyethylmethacrylate (HEMA) and methacrylic acid (MA), have awater content of 58%, and a modulus of 0.3 MPa. Narafilcon A lensescontain silicone, have a water content of 46%, and a modulus of 0.66MPa.

A saline commercial lens cleaning fluid (Sensitive Eyes® Plus salinesolution) was obtained from Bausch & Lomb, and is referred to herein as“B&L saline”.

Water used was purified by Barnsted NanoPure systems to a resistance of18.2 MΩ-cm resistance with total organic content levels of less thanapproximately 1 part per billion.

Liposome Preparation (Multilamellar Vesicles):

Multilamellar vesicles (MLV) composed either ofdimyristoylphosphatidylcholine(1,2-dimyristoyl-sn-glycero-3-phosphocholine; DMPC) or of hydrogenatedsoy PC (HSPC) were prepared by hydrating the lipids at a temperature atleast 5° C. above the lipid melting point (T_(M)), followed bysonication, in phosphate buffer saline (PBS). Where MLV liposomes weremixed with hyaluronic acid (HA), the polymer solution (in PBS) wasprepared in advance, and after full dissolution of the HA, the solutionwas warmed to a temperature at least 5° C. above the lipid T_(M), andadded to the lipids, followed by stirring to mix.

Liposome Preparation (Small Unilamellar Vesicles):

Multilamellar vesicles (MLV) composed of dimyristoylphosphatidylcholine(DMPC) or hydrogenated soy PC (HSPC) were prepared by hydrating thelipids at a temperature above the lipid melting point (T_(M)), accordingto the procedures described hereinabove. In order to obtain smallunilamellar vesicles (SUV), the MLVs were downsized by stepwiseextrusion through polycarbonate membranes, starting with a 400 nm andending with 50 nm pore size membrane, using a Lipex 100 mL extrudersystem (Northern Lipids, Vancouver, Canada), at a temperature above thelipid T_(M).

Where SUV liposomes were mixed with a polymer, the polymer solution (inPBS) was prepared in advance, and after full dissolution of the polymer,the polymer solution was added to the lipids, followed by stirring tomix for 2 hours.

Multilamellar vesicles and small unilamellar vesicles composed of otherpure phosphatidylcholines, such as dipalmitoylphosphatidylcholine(DPPC), dilauroylphosphatidylcholine (DLPC) and/ordistearoylphosphatidylcholine (DSPC), according to the proceduresdescribed hereinabove.

Friction Measurements:

Lenses were removed from their container, where they had been immersedin a phosphate buffer saline (PBS) solution, and were rinsed using PBS.The lenses were then immersed for 2 days in a PBS solution of liposomesand/or a polar polymer (hyaluronic acid (HA), polyvinylpyrrolidone (PVP)or polyethylene oxide (PEO)), or in PBS alone (as a control).

Prior to measurements in the tribometer, in all samples (including thecontrols), the lenses were thoroughly rinsed by a stream of B&L salineor PBS. The lenses were then mounted on the tribometer holder andfriction forces measured while sliding against a glass surface andimmersed in B&L saline or PBS.

Friction tests were performed with a UMT model tribometer (Bruker).Contact lenses were mounted on a cornea-mimicking holder, which has atypical geometry resembling the human cornea, as shown in FIGS. 1A and1B. The contact lens was then positioned opposite a glass plate andimmersed in B&L saline or PBS during the measurement. The normal loadsused were 3 grams, 5 grams, 10 grams and 40 grams.

The glass substrates used were thin 24 mm×24 mm cover-glasses (KnittelGlaser, Germany). They were removed from their pack (edge-handled withlatex gloves throughout), and glued into a standard 35 mm diameterpolystyrene Petri dish using Devcon® 5 Minute® 2-component epoxy. Justprior to the friction measurements, the upper glass surface was wipedwith an ethanol-moistened Kimwipes® tissue, then rinsed in de-ionizedwater to remove any ethanol traces, and the Petri dish then filled withthe B&L saline or PBS.

The friction coefficient was calculated by dividing the measured lateralforce during sliding by the applied normal force. Friction coefficientvalues are those of kinetic friction, which is related to the forces inthe system that are measured when there is a sliding motion of thecontact lens on the opposing glass surface. Parameters were as follows:sliding velocity 1 mm per second, frequency 1 Hz, and dwell time of 5seconds prior to initiation of motion. Experiments were conducted at atemperature of 36±0.5° C. or 37±1° C.

Each friction coefficient value (μ) is an average of frictionmeasurements for at least 5 different etafilcon A (HEMA/MA) lenses, orfor at least 3 different narafilcon A (silicone) lenses, for eachimmersion condition. Moreover, each friction measurement is an averageover 180 cycles for each of 2 to 3 different contact position on theglass surface. The same glass surface was used for one entire set ofexperiments for a given lens type, and the order of measurements was asfollows: first, saline controls; then a lens that had been immersed inHA; then a lens that had been immersed in HSPC; then a lens that hadbeen immersed in HSPC+HA; then a lens that had been immersed in DMPC;then a lens that had been immersed in DMPC+HA. Between each differentlens the B&L saline or PBS immersing the lens/substrate system wasreplaced by fresh B&L saline or PBS, respectively. The glass surface wasthen changed, and the measurements repeated (5 times for etafilcon A and3 times for narafilcon A).

In one case, following the full set of measurements with a given glasssubstrate, the measurement for the (HSPC+HA)-immersed lens on the samesubstrate was repeated, and the earlier measured value (for the same(HSPC+HA)-immersed lens) was reproduced.

The mean pressure P over the contact area A was determined according tothe equation: P=F_(N)/A, where FN is the applied normal load and, fromHertzian contact mechanics [Johnson, K. L., Contact Mechanics 2004,London: Cambridge University Press], A=π(RF_(N)/K)^(2/3), where R is theradius of the rigid cornea-mimicking holder and K is the Young's modulusof the contact lens.

Dynamic Light Scattering (DLS):

Dynamic light scattering (DLS) measurements of the various suspensionswere determined using a ZetaSizer μ V apparatus (Malvern Instruments).

Results

Dynamic light scattering (DLS) measurements showed that HA in PBS had ahydrodynamic diameter of 135±20 nm. For the MLV HSPC and DMPC liposomesin PBS solution, DLS measurements yielded diameters of 3±1.5 μm and1.4±0.7 μm, respectively.

DLS measurements of the MLV's HSPC and DMPC liposomes mixtures with HAindicated diameters of 2.5±1.5 μm and 2.8±1.5 μm, respectively.

Friction coefficients were measured for Etafilcon A and Narafilcon Alenses, which served as exemplary hydrogel surfaces, in B&L salineenvironment at 36±0.5° C., according to procedures describedhereinabove, either following removal of the lens from the blister-packand rinsing in B&L saline (labeled ‘saline’ in the figure legends), orfollowing immersion in PBS solutions containing the tested liposomes(HSPC and DMPC MLVs at a concentration of 45 mM) and/or 1 MDa HA (1 M;0.2 mg/ml), and rinsing in B&L saline.

The applied loads (L) were 5 grams, 10 grams or 40 grams, and thecorresponding mean pressures P (in Atm units) are presented in FIGS. 2and 3 , respectively as L/P.

As shown in FIGS. 2 and 3 , the sliding friction coefficients μ ofhydrogel lenses that were only rinsed in B&L saline following removalfrom their blister-pack, and then slid across a glass slide immersed inB&L saline, was in the range 0.08±0.04 for HEMA hydrogel (Etafilcon A)and 0.2±0.1 for silicone hydrogel (Narafilcon A). These values areconsidered as the baseline control relative to the values obtained withother solutions, and are designated herein asp.

As further shown in FIGS. 2 and 3 , following immersion in HA solution,the sliding friction coefficient μ decreased relative to the baselinevalue μ₀, by 30% and 50%, for the Etafilcon A and the Narafilcon Ahydrogel lenses, respectively.

Following immersion in liposome solutions, a significant reduction inthe sliding friction coefficient μ relative to μ₀ was generally noted,ranging between 25% to about 75% for the HSPC liposomes and between 65%to 92% for the DMPC liposomes.

Following immersion in the HA/liposome mixtures, substantially higherreduction in sliding friction coefficients μ relative to μ₀ wereinvariably observed, ranging from about 2-fold reduction for Etafilcon(HEMA) immersed in HA+HSPC to more-than-10-fold reduction for Narafilcon(silicone) immersed in HA+DMPC.

In some cases, the friction coefficients were somewhat lower at thehigher loads.

These results present a synergistic effect of a solution containing bothHA and liposomes. It is to be understood that in sliding frictioncoefficient, when two or more lubricants are measured alone and incombination, it is expected that the combination would result inaveraged values of the friction coefficient. However, surprisingly, asolution containing HA and the liposomes resulted in frictioncoefficient values which were substantially lower than the frictioncoefficient values obtained for either component alone, thusdemonstrating a synergistic effect.

HA is known not to be a good boundary lubricant [Seror et al.,Biomacromolecules, 13(11):3823-3832, (2012)]; Benz et al. Journal ofBiomedical Materials Research Part A, 2004. 71A:6-15], although viscoussolutions of HA, similarly to other viscous solutions, have beenconsidered to act as non-boundary lubricants [Doughty, Contact Lens andAnterior Eye 1999, 22:116-126].

It is noted that all measurements were performed following 2-dayimmersion of the hydrogel lenses in the tested solutions and asubsequent thorough rinse in a stream of B&L saline, such thatsubsequent measurements were made in B&L saline alone. It is thereforeassumed that there was no trace of free HA or liposomes in the liquidsurrounding the hydrogel lenses in the tribometer.

Friction coefficients were then measured for Etafilcon A and NarafilconA hydrogel lenses in a PBS environment at 37±1° C., according toprocedures described hereinabove, following immersion in PBS solutionscontaining the HSPC and DMPC SUVs (at a concentration of 10 mM) and/or apolar polymer (1.5 MDa hyaluronic acid (HA), polyvinylpyrrolidone (PVP)or polyethylene oxide (PEO) at a concentration of 0.2 mg/ml), or in PBSalone (as a control).

Dynamic light scattering (DLS) measurements showed that HSPC SUVs had adiameter of ˜100 nm, and DMPC SUVs had a diameter of ˜72 nm.

The applied loads (L) were 3 grams or 10 grams, and the correspondingmean pressures (P) are presented in FIGS. 4-7 , respectively as L (ingrams)/P (in Atm units).

As shown in FIGS. 4-7 , and in Table 1 below, immersion in DMPC (FIGS. 4and 6 ) or HSPC (FIGS. 5 and 7 ) liposome solutions resulted in asignificant reduction in the sliding friction coefficient μ of EtafilconA (FIGS. 4 and 5 ) and Narafilcon A (FIGS. 6 and 7 ) hydrogel lensesrelative to hydrogel lenses immersed in PBS, in accordance with theresults described in Example 1.

As further shown in FIGS. 4-7 and in Table 1, immersion inpolymer/liposome mixtures generally resulted in substantially higherreduction in sliding friction coefficients μ than did immersion inpolymer solution or liposome solution, especially at a load of 10 grams.

It is noted that all measurements were performed following 2-dayimmersion of the hydrogel lenses in the tested solutions and asubsequent thorough rinse in a stream of PBS, such that subsequentmeasurements were made in PBS alone. It is therefore assumed that therewas no trace of free polymer or liposomes in the liquid surrounding thehydrogel lenses in the tribometer. Thus, the results indicate aninteraction and possible attachment of the polymers to the surface ofthe hydrogel.

Without being bound by any particular theory, it is believed thatresults at a load of 10 grams are more representative of long-termlubrication effects than are results at a load of 3 grams.

As shown in FIG. 4 and in Table 1, PVP/DMPC liposome and PEO/DMPCliposome mixtures resulted in a reduction of 50% or more in the frictioncoefficients of Etafilcon A hydrogel lenses in comparison with DMPCliposomes alone at a load of 10 grams.

As shown in FIG. 5 , PVP/HSPC liposome and HA/HSPC liposome mixturesresulted in a reduction of 25-30% in the friction coefficients ofEtafilcon A hydrogel lenses in comparison with HSPC liposomes alone at aload of 10 grams.

As further shown in FIGS. 4 and 5 , the abovementioned polymer/liposomemixtures resulted in a reduction of about 90% or more in the frictioncoefficients of Etafilcon A hydrogel lenses in comparison with PBS orpolymer solutions.

As shown in FIG. 6 , PVP/DMPC liposome and HA/DMPC liposome mixturesresulted in a reduction of 22-40% in the friction coefficients ofNarafilcon A hydrogel lenses in comparison with DMPC liposomes alone,and a reduction of 50-72% in comparison with PBS or the respectivepolymer solutions, at a load of 10 grams. As shown in FIG. 7 , PEO/HSPCliposome, PVP/HSPC liposome and

HA/HSPC liposome mixtures resulted in a reduction of 40-54% in thefriction coefficients of Narafilcon A hydrogel lenses in comparison withHSPC liposomes alone, and a reduction of 60-91% in comparison with PBSor the respective polymer solutions, at a load of 10 grams.

TABLE 1 Friction coefficients of Etafilcon A and Narafilcon A hydrogelcontact lenses under different loads and mean pressures, followingimmersion in PBS solution with or without liposomes and/or a polarpolymer (hyaluronic acid (HA), polyvinylpyrrolidone (PVP) orpolyethylene oxide (PEO)) Mean Polymer in PBS Solution Load (Atm)Liposome No Hydrogel (grams) Pressure type polymer HA PVP PEO Etafilcon 3 0.1  No 0.21 ± 0.055 ± 0.02 ± 0.2 ± A liposomes 0.07 0.01 0.005 0.02DMPC 0.015 ± 0.012 ± 0.01 ± 0.009 ± liposomes 0.005 0.006 0.005 0.003HSPC 0.016 ± 0.015 ± 0.011 ± N.D. liposomes 0.007 0.005 0.003 10 0.16 No0.28 ± 0.31 ± 0.11 ± 0.45 ± liposomes 0.055 0.1 0.03 0.05 DMPC 0.024 ±0.024 ± 0.012 ± 0.009 ± liposomes 0.007 0.008 0.004 0.003 HSPC 0.024 ±0.017 ± 0.018 ± N.D. liposomes 0.009 0.005 0.003 Narafilcon  3 0.18 No0.051 ± 0.025 ± 0.02 ± 0.09 ± A liposomes 0.015 0.005 0.01 0.03 DMPC0.015 ± 0.016 ± 0.01 ± N.D. liposomes 0.005 0.005 0.0035 HSPC 0.02 ±0.018 ± 0.013 ± 0.012 ± liposomes 0.004 0.006 0.004 0.004 10 0.26 No0.067 ± 0.05 ± 0.054 ± 0.12 ± liposomes 0.025 0.017 0.017 0.028 DMPC0.032 ± 0.025 ± 0.019 ± N.D. liposomes 0.005 0.005 0.007 HSPC 0.033 ±0.02 ± 0.016 ± 0.015 ± liposomes 0.008 0.01 0.008 0.006 N.D. = notdetermined

As further shown in FIGS. 4-7 , mixtures of the non-ionic polar polymersPVP and PEO with liposomes resulted in at least as great a reduction insliding friction coefficients μ as did immersion in mixtures of theionic polymer hyaluronic acid with liposomes.

These results indicate that solutions containing ionic or non-ionicwater-soluble polymers and the liposomes resulted in frictioncoefficient values which were substantially lower than the frictioncoefficient values obtained for either component alone, thusdemonstrating a synergistic effect.

These results further indicate that SUV liposomes are highly effectiveat reducing friction coefficients (as are MLV liposomes described inExample 1) in combination the polar polymers.

As further shown in FIG. 4 , a mixture of PEO and DMPC SUVs wasparticularly effective at reducing sliding friction coefficients ofEtafilcon A hydrogel, whereas PEO alone had no effect on the slidingfriction coefficient at a relatively low load (3 grams), and resulted inan increased sliding friction coefficient at a higher load (10 grams).

Similarly, as shown in FIG. 7 , a mixture of PEO and HSPC SUVs wasparticularly effective at reducing sliding friction coefficients ofNarafilcon A hydrogel, whereas PEO alone resulted in increased slidingfriction coefficients.

These results surprisingly indicate a particularly strong synergy (atreducing friction coefficients) between PEO (which is not effective atreducing friction coefficients by itself) and liposomes of differenttypes, and on different surfaces.

Some Non-Limiting Mechanistic Insights:

Without being bound by any particular theory, the following provides atentative explanation of the results presented above.

The reduction in the friction coefficient upon immersion in polarpolymer solution and a subsequent rinse in B&L saline or PBS may beregarded as evidence of an interaction and possible attachment of thepolar polymer to the surface of the hydrogel.

The higher reduction (relative to saline and to polar polymer solutions)in the friction coefficient upon immersion in liposomes solution and asubsequent rinse in B&L saline or PBS may be regarded as evidence ofcoverage of the hydrogel surface.

PC liposomes are well known to act as efficient boundary lubricants,hence the (generally observed) reduction in μ relative top.

It is assumed that at relatively low pressures the DMPC lipids providebetter lubrication than the HSPC, possibly because that at 36-37° C.,the DMPC are in their liquid disordered (LD) phase (T_(M)(DMPC)=24° C.)and hence are more highly hydrated than the HSPC, which at 36-37° C. isin its solid ordered (SO) phase (T_(M)(HSPC)=53° C.), whereas at highertemperatures (including those used in previous studies), the situationis reversed, and HSPC liposomes are the better lubricants since theirbilayers are more robust than the DMPC ones [Goldberg, R., et al.,Advanced Materials, 2011, 23:3517-3521; Sorkin, R., et al.,Biomaterials, 2013. 34:5465-5475]. This may explain the differences inthe relative efficacy of DMPC and HSPC liposomes under loads of 3 gramsand 10 grams, for example, as shown in Table 1.

When the hydrogel lenses are immersed in a mixture of the liposomes andpolar polymer, polar polymer adsorbs on the hydrogels and, in thissurface-attached form, complexes with the lipids to form highlylubricating boundary layers.

These findings are further supported by the studies described inExamples 2 and 3 below.

These findings are also qualitatively consistent with the somewhatweaker effect that HA has either on its own or, synergistically, withthe liposomes, when Etafilcon hydrogels (HEMA+MA groups) are usedrelative to Narafilcon hydrogels (silicone).

The Etafilcon hydrogel is slightly negatively charged due to themethacrylic acid (MA) groups, whereby the Narafilcon is uncharged. HAexhibits both negative charge and hydrophobicity. It is thereforeassumed that while HA may interact via hydrophobic and electrostaticinteractions, it adheres more weakly to negatively-charged surfaces suchas HEMA. This lower absorbance of HA on the Etafilcon accounts for theweaker reduction in friction for Etafilcon vs. Narafilcon, both when HAalone is used, and when it is used together with liposomes in theimmersing solutions, thus indicating a role for HA absorbance to thehydrogel surface in reducing friction coefficient and increasinglubricity.

Example 2 Phosphatidylcholine Liposomes-Hyaluronic Acid SurfaceComplexes Introductory Comments

The origin of the boundary lubrication in mammalian synovial joints hasbeen studied for decades but a generally accepted consensus is stillelusive. HA is one of the main macromolecules composing the cartilagetissue and, anchored by entanglements within the collagen network or bylubricin within the superficial zone (SZ), is present also at its outerinterface with the synovial cavity, as schematically indicated inBackground art FIG. 4 . Phospholipids are present both in the synovialfluid (SF) and in the cartilage superficial zone, and indeed DPPC hasbeen suggested as being among the most abundant phospholipids both in SFand in articular cartilage.

In addition to lubricin, both phospholipids and HA have long beenimplicated in cartilage boundary lubrication. Separately, theinteractions between HA and DPPC lipids have been studied independentlyby several groups.

Herein, a combined effect of HA and phospholipids on the slidingfriction coefficient of surfaces, at pressures mimicking those ofarticular joints, has been studied.

Materials and Methods

Materials:

Water for the SFB experiments and the AFM imaging under water waspurified with a Barnstead water purification system (Barnstead NANOpureDiamond, resistivity=18.2MΩ, total organic content (TOC)<1 ppb.

Ruby Muscovite mica grade 1 was obtained from S & J Trading, Inc.,

NY was utilized for the SFB experiments and for the AFM and Cryo-SEMimaging.

Avidin from egg white (A9275) was obtained from Sigma Aldrich, Israel.

Potassium Nitrate salt (purity >99.99%) was obtained from Merck;

DPPC lipids were obtained from Lipoid GmbH;

medical-grade HA (0.5 to 1.5 MDa) for the biotinylation was obtainedfrom Genzyme;

non-biotinylated HA (1 MDa) was obtained from Lifecore Biomedical;

biotin-LC-hydrazide and EDAC were obtained from Pierce and Warriner,Chester, UK.

Biotinylation of HA:

The biotinylation of HA was performed as described in detail in Mahoney,D. J., Blundell, C. D. & Day, A. J. [Journal of Biological Chemistry2001, 276:22764-22771] and Seror et al. [Biomacromolecules, 2011,12(10):3432-3443]. In brief, 5 mg of HA was dissolved overnight in 0.1MMES, pH 5.5, at a concentration of 5 mg/ml. 13 μl of a solution of 25mg/ml EDAC in 0.1M MES, pH 5.5, were then added, followed by addition of20 μl of 50 mM biotin-LC-hydrazide in dimethyl sulfoxide. The reactionmixture was mixed by rotation at room temperature overnight, and wasthereafter dialyzed extensively against water and particulate materialremoved by centrifugation (12,000×g for 1 minute).

The concentration of the bHA was determined using themetahydroxybiphenyl reaction [Blumenkrantz, N. & Asboe-han, G. (1973)Analytical Biochemistry 54, 484-489] relative to standards made from HAdried in vacuo over cobalt chloride. The bHA (in 0.02% (w/v) NaAzide)was stored at 4° C.

Liposomes Preparation:

Multilamellar vesicles (MLVs) were prepared by hydrating DPPC at 70-75°C. (above its solid-ordered-to-liquid-disordered transition temperatureT_(M)=41° C.). MLVs were then downsized to form small unilamellarvesicles (SUVs), about 80 nm in diameter, by stepwise extrusion throughpolycarbonate membranes starting with a 400 nm and ending with 50nm-pore-size membrane, using a Lipex 100 mL extruder system (NorthernLipids, Vancouver, Canada). The size distribution of the SUV wasdetermined by dynamic light scattering.

Atomic Force Microscopy (AFM):

Avidin-bHA-DPPC-coated mica: Freshly cleaved mica was glued on a Petridish and soaked in 0.01 mg/ml avidin aqueous solution for about 30minutes and then rinsed in water for about 1-2 minutes. The sample wasthen covered with 49 μg/ml bHA solution and kept in a humiditycontrolled chamber for several hours. After rinsing the sample withexcess of water, the Petri dish was filled with 5 ml of water, to which0.2 ml of 15 mM suspension of DPPC liposomes (SUVs, prepared asdescribed hereinabove) was added. After overnight adsorption the sampleswere rinsed in water and scanned with an Asylum MFP3D under pure waterusing a Veeco-SNL tip (radius of about 2 nm).

HA-DPPC liposomes mixed in the bulk: 1 mg/ml HA and 1 mg/ml DPPCliposomes in the form of SUV liposomes (prepared as describedhereinabove) were stirred together in the dark for 24-48 hours at atemperature of about 60-70° C. (above the liposomes' T_(M)), inaccordance with a published protocol [Pasquali-Ronchetti, I., Quaglino,D., Mori, G. & Bacchelli, B. (1997) JOURNAL OF STRUCTURAL BIOLOGY 120,1-10]. A freshly cleaved mica surface, previously glued on a Petri dish,was covered with the HA-DPPC solution (after cooling to roomtemperature) and kept overnight in a humidity controlled chamber. Thesample was then rinsed with water, while avoiding exposure to air, andscanned as described above.

AFM samples of both configurations were identical to the surfaces usedin the SFB measurements.

SFB Measurement Procedure:

The Surface Force Balance (SFB) measurements were performed aspreviously described [Klein, J. & Kumacheva, E. (1998) Journal ofChemical Physics 108, 6996-7009; Klein, J. (1983) Journal of theChemical Society-Faraday Transactions I 79, 99; Raviv, U. & Klein, J.(2002) Science 297, 1540-1543], and as schematically illustrated in FIG.10C, involving measurement of normal and shear interactions betweenmolecularly smooth sheets of mica at separation D (whose absolute valueis measured to ±2-3 Å).

Avidin-bHA-DPPC-coated mica: HA was attached to the substrate asfollows: following calibration at bare-mica/bare-mica contact, thesurfaces were soaked in 0.01 mg/ml avidin aqueous solution for about 30minutes and then rinsed in water for 1-2 minutes. Attachment of HA wasachieved by interacting lightly biotinylated HA (bHA) with the avidin onthe mica via the avidin-biotin interaction (and, partly, viaelectrostatic interactions between the negative HA and the positiveavidin), as previously described [Seror et al. (2011) Biomacromolecules12, 3432-3443; Seror et al. (2012) Biomacromolecules 13, 3823-3832].

Normal and shear interactions between the avidin-bearing and, followingthat, between avidin-HA-bearing surfaces were generally measured ascontrols to ensure the integrity of the surface layers prior tointroduction of the liposomes. Samples where contaminant-free attachmentof HA on the mica was indicated were used in the next stage.

The HA-coated mica surfaces on their lenses were immersed overnight in10 ml of pure water into which 400 μl of 15 mM of a suspension DPPCliposomes (SUVs, prepared as described hereinabove) was added, and thenrinsed in 400 ml of pure water and remounted in the SFB as close aspossible to their original position.

Normal and shear interactions were then measured between theavidin-bHA-DPPC bearing surfaces. Finally, water was substituted with0.15 M KNO₃ solution and normal and shear interactions were measuredagain.

The results are based on 5 different experiments and 2 to 4 differentcontact positions on each experiment. The mean pressure P was evaluatedas P=F_(n)/A, where F_(n) is the applied normal force; the contact areaA=πa² or nab where a and b are principal radii of the circular (a=b) orelliptical contact area arising from elastic flattening of the gluebeneath the mica sheets (measured directly from the flattening of theinterference fringes [See, Chen et al. (2009) Science 323, 1698-1701;Goldberg et al. (2011) Advanced Materials 23, 3517-3521; Sorkin et al.(2013) Biomaterials 34, 5465-5475]. An uncertainty of ±(15-20) % in Pdue to uncertainties of order 10% in the measured radii, was estimated.

Results

AFM Characterization:

FIG. 9A (main) presents an AFM micrograph of a mica surface afterovernight incubation in a solution of DPPC-SUVs and HA, which waspreviously mixed for 48 hours at 60° C. in the dark, followed by rinsingin water, demonstrating that the SUVs are adsorbed in a close-packedconfiguration on the surface. Inset (i) in FIG. 9A shows the micasurface after overnight incubation in a solution of DPPC-SUVs alone,which previously mixed for 48 hours at 60° C. in the dark, followed byrinsing in water. Inset (ii) in FIG. 9A is CRYO-SEM figure, taken fromSorkin et al. [(2013) Biomaterials 34, 5465-5475], and showing part of acryo-SEM micrograph of mica following incubation in a (HA-free) DPPC-SUVdispersion in water at room temperature.

As shown in FIG. 9A, there is little difference between the threesurface configurations, demonstrating that any interaction of HA withthe DPPC-SUVs in the bulk dispersion leads to little change in theirinteractions with the mica. It is to be noted that due to compression bythe tip, the AFM measurements indicate vesicle dimensions normal to thesurfaces that are likely to be considerably compressed relative to theirunperturbed thickness (see, Goldberg et al. (2011) supra).

FIG. 9B presents AFM micrographs of an avidin-bHA-bearing-mica surfacefollowing incubation in (HA-free) DPPC-SUV dispersion. As shown therein,a very different structure compared to those presented in FIG. 9A isobserved, whereby the surface is densely covered with elongated,beads-on-string-like structures of around 6-10 nm thickness; two suchcontours are indicated as a guide to the eye. The substantial differencebetween these structures and the adsorbed vesicles when HA is in thebulk rather than on the surface is highlighted by the inset of FIG. 9B,which shows, on the same scale, one such vesicle taken from the AFMmicrograph in FIG. 9A, for comparison. Thus, it is clearly shown thatthe vesicles, which originally had a DLS-determined radius of about 44±5nm (or about 30-45 nm as revealed by cryo-SEM when adsorbed on the baremica surface from the HA-DPPC mixture, as shown in FIG. 9A) haveruptured to form complexes with the surface-attached HA as shown in FIG.9B.

It is to be noted that the height of the HA-DPPC complexes appearssmaller (2.5-3 nm) than their width (6-10 nm). This may be attributedpartly to compression by the AFM tip (which may indent the bilayers),and also to the fact that the HA chains are attached to avidin groups(height of about 4-5 nm on the mica [see, Seror et al. (2011)Biomacromolecules 12, 3432-3443] and so may be compressed into the gapstherebetween.

FIG. 9C presents a schematic illustration of the obtained complexes,which, without being bound by any particular theory, are assumed to becomposed of HA chains, whose uncomplexed thickness is around 0.3-1.5 nm[see, Jacoboni, et al. (1999) Journal of Structural Biology 126, 52-58],enclosed by DPPC lipid monolayers or bilayers (about 5 nm thickness fora bilayer).

Surface Interactions:

Normal Surface Forces:

Using the SFB, both normal and shear interaction profiles, F_(n)(D) andF_(s)(D) respectively, were measured (see, FIG. 10C) between micasurfaces bearing DPPC attached from incubating solutions containing HA,and forming the two configurations presented in FIGS. 9A and 9B anddiscussed hereinabove.

FIG. 10A presents the normalized interactions between two HA-DPPCsurface complexes (shown in FIG. 9B). The range of interactions varieswithin about ±10% of the mean range between different experiments, butless than that within different contact points of a given experiment. Asshown in FIG. 10A, a common trend is recognizable in the majority of theprofiles. Generally, first approaches are longer ranged with repulsionstarting already at surface separation around D of about 150-250 nm(full symbols in FIG. 10A), while in second or subsequent approaches(crossed symbols in FIG. 10A) the surfaces start to repel each otheronly at a separation around D of about 60-120 nm.

As shown in FIG. 10B, first profiles often present a kink at aseparation of between 60-100 nm (that is, the forces remain roughlyconstant over this separation range), after which they continue toincrease monotonically. The kink may be interpreted as squeezing out ofthe residual liposomes by the compressing surfaces.

Receding force profiles (open symbols in FIG. 10A) have a similar trendto second approaching profiles (crossed symbols in FIG. 10A). Thesefeatures may readily be attributed to residual loose, weakly attachedvesicles overlaying the firmly-attached HA-DPPC surface complex seen inFIG. 9A, arising from inadequate rinsing following incubation. Theseloose vesicles may be removed by the compression and the shearing motionduring the approach, leading to shorter interaction range at separationand subsequent approaches.

The final separation at high pressures reached in first and subsequentapproaches is very similar: D=22±3 nm and 23±3 nm, respectively, or some11 nm/surface. This may be attributed to the thickness of the avidin(about 4 nm), covered by HA (about 0.9 nm) complexed with a DPPC bilayer(about 4-5 nm), which account for some 9-10 nm before consideration ofany chain overlap on the surface (see, e.g., FIG. 9C).

Red full and crossed symbols in FIG. 10A are respectively first andsecond approaches of the avidin-bHA-DPPC bearing surfaces in 0.15 M saltsolution. The shorter onset range of the repulsion in salt solution(about 100 nm) relative to pure water (150-200 nm) is attributed toremoval of residual intact liposomes, due both to shear in the purewater prior to adding salt, and to the effect of replacing pure water bythe salt solution which is effectively an additional rinsing stage.

Lateral/Frictional Forces:

FIGS. 11A and 12 present the shear force vs. time traces, F_(s)(t),between mica surfaces, for the configuration where the mica surfaces arecoated with the HA-DPPC complexes (shown in FIGS. 9B and 9C), recordeddirectly from the SFB, across water and across 0.15M KNO₃, respectively.In FIGS. 11A and 12 , the top saw tooth traces represent the back andforth motion of the upper surface as a function of time, while tracesbelow are the corresponding shear forces between the surfaces recordedat different mean pressures P (arising from different loads F_(n)) and Dvalues, at given contact points. The plateaus in the shear force tracescorrespond to sliding of the surfaces.

FIG. 11B shows variation of the shear force under high compression(P=161 atm, D=20 nm) over some 3 orders of magnitude in the slidingvelocity v_(s), indicating little change in F_(s), a signature ofboundary lubrication.

FIG. 11C shows that the surfaces are robust to prolonged sliding at highP values, as F_(s) does not increase over time (for periods up to anhour), and may even decrease. This decrease may be attributed torearrangement under sliding of the surface-attached complexes to a lessdissipative orientation.

FIG. 13A presents a summary of the shear force vs. load results andshows an initial rapid rise in the friction at lower loads (andpressures). This phenomenon is attributed to the viscous dissipationarising from shear of the loosely-attached liposomes on top of theHA-DPPC complex attached to the surface, once the surfaces arecompressed to the range of steric repulsion at D<ca. 100 nm (see, FIGS.10A-10C). On shear at progressively higher pressures, the looselyattached vesicles are, as noted above, squeezed out of the contactregion, as indicated by the lower shear forces on a second approach at agiven contact point (prior to reaching the ‘hard wall’ separation). Atthe highest compressions—of order 50-100 atm or higher—the surfacesreach their limiting separation of 22±2 nm, corresponding to F_(n)>ca.10 mN (see F_(n)(D) profile in FIGS. 10A-10C). At these compressions theHA-DPPC complexes, firmly attached to each surface, are sliding directlypast each other, and the effective friction coefficient μ=F_(s)/F_(n),while showing some scatter (see shaded region in FIG. 13A), has a valueμ of about (1.5±1)×10⁻³ (taken over all experiments and contact points).

FIG. 13B presents a comparison of the F_(s) vs. F_(n) variation betweenmica surfaces with an avidin-bHA layer but in the absence of any addedliposomes, highlighting the orders of magnitude decrease in frictiononce the surface attached HA is complexed with DPPC.

The data with added salt (FIG. 13A, red symbols) shows a similar trend,while the friction coefficient is slightly higher. This is attributed toa reduced hydration level of the phosphocholine headgroups in thepresence of high salt and consequently a less efficient hydrationlubrication mechanism (as previously described; see, e.g., Chen et al.(2009) Science 323, 1698-1701). At the higher loads (F_(n)>ca. 2-3 mN,corresponding to D of about 22 nm, P>ca. 50 atm) where thesurface-attached HA-DPPC complexes are sliding directly past each other,the friction coefficient μ is about (7±1)×10⁻³.

Overall, the findings described herein indicate that DPPC lipids,introduced into the system as liposomes, complex with HA when HA isattached to the interacting surfaces, and these HA-DPPC complexes resultin robust boundary layers that provide excellent lubrication (down tofriction coefficient μ of about 10⁻³) up to mean contact pressures ofabout 200 Atm. The exceptional lubrication obtained with such complexessubstantially exceeds the lubrication obtained when HA alone is attachedto the surfaces, as seen in FIG. 13B.

When hydrogenated soy phosphatidylcholine (HSPC) SUVs rather than DPPCSUVs were used, the obtained results (not shown) were results to thoseshown in FIGS. 9A-9B, 10A-10B, 11A-11C, 12 and 13A-13B. This issuggestive, as HSPC, while not native to cartilage, is a saturateddiacyl PC, with predominantly 18:0 (˜85%) and 16:0 (˜15%) fatty acyltails; and such saturated 16:0 and 18:0 tails comprise some 30% of thePCs at the cartilage surface.

It is noted that HA is negatively charged, and thus the dipolarphosphocholine head-groups of phosphatidylcholine lipids (such as DPPC)presumably experience a dipole-charge attraction to the polysaccharide.Since the surface exposed by the HA-DPPC complexes must be hydrophilic,it is assumed that the structure of these complexes is either a bilayer,where the lipid headgroup attaches to the negative charge on the HA, ora DPPC monolayer where the acyl tails of the lipid attach viahydrophobic interactions to the hydrophobic patches on the HA chains(about 8 CH unit per disaccharide [see, Laurent, T. (1989) CibaFoundation Symposia 143, 1-5; Scott, J. E. (1989) Ciba FoundationSymposia 143, 6-20].

HA-DPPC complexes that have been imaged with negative-staining[Pasquali-Ronchetti, et al., Journal of Structural Biology 1997,120:1-10] show that HA complexes with DPPC, augmenting the HA contourthickness.

The width of the HA-DPPC complexes (6 to 10 nm) measured as describedherein (see, FIG. 9B) supports either of the two possible configurationsdescribed above, i.e. an HA chain surrounded by two bilayers or twomonolayers or a combination of the two.

The observations described herein can be regarded as indicating thatsurface-attached HA chains are coated with DPPC layers exposing theirhighly hydrated phosphocholine headgroups; thus rendering the micasurfaces coated with such layers invariably wet when withdrawn fromwater.

When the same DPPC-SUVs are well-mixed with HA in bulk solution ratherthan attached to the surface, HA does not appear to disrupt thevesicles, as a close packed liposome surface-layer is obtained, which issimilar to that obtained in the absence of HA (see, FIG. 9A).

Significantly, a very low friction coefficient was measured between micasurfaces having the HA-DPPC complexes at high pressures. At pressures ofabout 50 Atm (comparable to mean pressures in major joints), anyresidual loose vesicles have been squeezed out of the gap, and thepolysaccharide-lipid complexes are in direct contact as they slide pasteach other. The robustness to sliding at high compression isdemonstrated by the constancy (or even decrease) of the friction,following an hour of continuous sliding (see, FIG. 11C), and the weakvariation of the friction with sliding velocity over 3 orders ofmagnitude in the latter, demonstrated in FIG. 11B, provides a furtherindication of boundary lubrication. The low friction between the HA-DPPCcomplexes, as they slide past each other, is presumably attributed tothe hydration lubrication mechanism, which, further presumably, arisesfrom the fluid nature of tenaciously-held hydration layers, particularlyfor the case of the highly-hydrated phosphocholine groups exposed bythese surface structures.

The results presented herein are brought, inter alia, as an explanationof the mode of action of the HA and liposomes solutions described inExample 1 in the context of contact lens. That is, HA attaches to thesurface of the lenses (e.g., by adsorption), and the liposomes thencomplex with this surface-attached HA to form the robust,highly-lubricating boundary layer.

These results further demonstrate that using HA and liposomes forproviding lubricity is efficiently performed by attaching or tetheringHA to the surface to be lubricated (e.g., a surface which does notadsorb HA effectively), such that the liposomes interact with thetethered HA to form the HA-lipid complexes comprising thehighly-lubricating boundary layer.

These results further provide a clearer understanding of the boundarylubrication of joints. The articular cartilage collagen network is knownto be permeated with HA, which in time diffuses through the outercartilage surface into the synovial cavity [Klein, J. (2006) Proceedingsof the Institution of Mechanical Engineers Part J—Journal of EngineeringTribology 220, 691-710]. During its transport through this interface theHA may still be slowed down by entanglements within the cartilage SZ(superficial zone) (as indicated in Background art FIG. 8 ), or, morelikely, be held at the surface by its interactions with SZ lubricin, andwill complex with the phosphocholines (PCs) that are ubiquitous both incartilage and in the SF. As shown herein, such complexed,surface-attached HA-PC structures can provide robust boundarylubrication with friction coefficients μ of about 10⁻³, mimicking thosein the major mammalian joints up to the highest pressures in suchjoints.

These findings indicate that HA, lubricin and phospholipids possibly acttogether to provide the remarkable lubrication of articulatingcartilage: Superficial zone lubricin is responsible, at least in part,for tethering HA at the cartilage surface; the surface-bound HA, asshown herein, in turn complexes with the cartilage/SF PCs; and theseboundary HA/PC complexes, acting via the hydration lubricationmechanism, provide the low friction that is the hallmark of healthysynovial joints, and further account for the natural replacement of theboundary layers as they wear away (since HA originating either in thecartilage chondrocytes or in the SF is continuously permeating anddiffusing through the cartilage space, or through the SF, to arrive atthe superficial zone and at the cartilage outer surface, where, held bythe SZ lubricins, it may complex with phospholipids to replenish theboundary layer).

These findings further suggest that efficient treatment of arthritic(e.g., osteoarthritic) joints can be performed by attaching hyaluronicacid to the surface of a joint (e.g., to cartilage) via a linkerdesigned to bind to hyaluronic acid and to collagen (e.g., collagen II),and administering liposomes. Such a linker optionally comprises acollagen-binding peptide (e.g., collagen II-binding peptide) for bindingto collagen in the joint (e.g., in cartilage at the articular surface)and a functional group or moiety which binds covalently and/ornon-covalently to the hyaluronic acid. Optionally, the linker comprisesa hyaluronic acid-binding peptide which binds to the hyaluronic acid.The hyaluronic acid is optionally administered with at least one linkerbound thereto, that is, in a form of a modified hyaluronic acid, forexample, modified hyaluronic acid comprising which comprises at leastone collagen-binding peptide (e.g., collagen II-binding peptide).

Example 3 Lubrication of Tendons by Hyaluronic Acid and Liposomes

Modified hyaluronic acid (HA) was modified by conjugating dopamine (DN)to the carboxylic acid groups of HA by a 1-ethyl-3-(3′-dimethylaminopropyl) carbodiimide (EDC) coupling reaction. The modified HA thuscomprised dihydroxyphenyl (catechol) groups, which have been reported topromote binding to organic surfaces (including amine-containingsurfaces) via covalent bond formation, as well as to inorganic surfacesvia strong non-covalent binding [Lee et al., PNAS 2006, 103:12999-13003;Brodie et al., Biomedical Materials 2011, 6:015014].

Briefly, 0.5 gram of HA was dissolved in 50 ml of PBS solution and thepH was adjusted to 5.5 using 1 N HCl solution. In the solution, 40 mg(0.05 mmol) of EDC and 94 mg (0.05 mmol) of dopamine hydrochloride wasadded and the pH of the reaction solution was maintained at 5.5 for 2hours with 1.0 N HCl and 1.0 N NaOH. Then, the solution was dialyzedagainst water for 2 days and was subsequently lyophilized, whichresulted in a white powder.

DN levels in the HA-DN conjugate were analyzed by ultraviolet (UV)spectrophotometry and nuclear magnetic resonance (NMR) analysis. For UVanalysis, a solution of 1 mg/ml in water was prepared. For 1H-NMR, thesample was dissolved in deuterated water (D₂O) for 3 hours atconcentrations of 2 mg/ml. The spectra were recorded at 298 K and 500MHz for 1H-NMR analysis. As shown by UV spectroscopy, an absorption bandat approximately 280 nm appeared for the HA-DN conjugate, which was notobserved for unmodified HA. Based on this band, it was determined thatthe concentration of dopamine units in the HA-DN solution was about0.075 mg/ml, which indicated that the degree of dopamine substitution inthe synthesized conjugate was about 19%. The catechol content (as amolar percentage, relative to repeating disaccharide units of HA) inHA-DN was determined by NMR analysis from the integral area ratiocalculation f=a/b, where a is the integral area of the peaks at around 7ppm, which corresponds to the amount of H in the aromatic rings ofgrafted catechol moieties, and b is the integral area of the peaks atabout 2.0 ppm, which represents the amount of H in the methylene ofpolymeric backbone. The degree of conjugated dopamine in the resultantpolymer was about 18% as determined by NMR analysis, which is consistentwith the result of UV analysis. Other batches of HA-DN were found tohave about 4% or 12% conjugated dopamine.

The friction coefficient characterizing friction between a chickentendon and its sheath under sliding condition was determined bypreparing tendon/sheath samples as depicted in FIGS. 14A-14E andmeasuring gliding resistance using a tribometer system depicted in FIG.15 .

The tendons were treated with solutions of hyaluronic acid (HA),hydrogenated soy phosphatidylcholine (HSPC) small unilamellar vesicles(SUVs), HSPC SUVs in combination with HA, or HSPC SUVs in combinationwith dopamine-functionalized HA (HA-DOPA, prepared as describedhereinabove), in phosphate buffer saline (PBS). Control tendons weretreated with PBS alone.

Before treatment, the friction force for all tendons was measured inPBS, under zero load force for calibration. Then the PBS was replacedwith the treatment solution, and the tendon was soaked in the treatmentsolution at 37° C. for 20 minutes. After 20 minutes, the treatmentsolution was replaced with PBS and the friction force between eachtendon and its sheath was measured.

As shown in FIGS. 16-18 , HA alone and HSPC liposomes alone each reducedthe friction coefficient of tendons, but the combination of HSPCliposomes with HA or dopamine-functionalized HA resulted in a reductionin the friction coefficient which was far more robust to repeated cyclesof friction than the reduction resulting from HA alone or HSPC liposomesalone, under loads of both 40 grams (FIGS. 16 and 18 ) and 80 grams(FIGS. 17 and 18 ). As further shown therein, dopamine-functionalized HAresulted in a considerably greater reduction in the friction coefficientthan did unmodified HA.

These results confirm that liposomes and polymers such as HAsynergistically reduce friction in physiological systems, and furtherindicate that polymers comprising functional groups which enhanceaffinity to a physiological surface such as a connective tissue are evenmore effective at reducing friction in such a system.

In order to assess the mechanism by which unmodified anddopamine-functionalized HA act in synergy with liposomes, HSPC SUVs werelabeled with the lipophilic fluorescent dye DiI(1,1′-dioctadecyl-3,3,3′3′-tetramethylindocarbocyanine) and the amountof lipids on tendon surfaces treated with HSPC alone or with unmodifiedand dopamine-functionalized HA was evaluated by fluorescencemeasurements.

As shown in FIGS. 19 and 20A-20C, unmodified and dopamine-functionalizedHA both increased binding of HSPC to tendon surfaces, withdopamine-functionalized HA being considerably more effective in thisrespect than unmodified HA. These results indicate that the synergisticeffect of liposomes and polymers such as HA is associated withenhancement by the polymer of the affinity of liposome lipids to asurface, and that polymers with enhanced affinity to the surface aremore effective at enhancing affinity of the lipids the surface.

Binding of liposome lipids to additional surfaces was evaluated byfluorescent measurements, using DiI-labeled liposomes as describedhereinabove, and a gelatin-methacrylate hydrogel surface. Unmodified HAwas used, as was HA functionalized with different levels of dopamine, 4%and 18% dopamine (relative to the number of repeating (disaccharide)units of HA).

As shown in FIG. 21 , unmodified and dopamine-functionalized HA bothincreased binding of HSPC to tendon surfaces, withdopamine-functionalization of HA enhancing the ability of HA to increaseHSPC binding in a manner which correlated to the level of dopaminegroups.

These results indicate that functionalized polymers such as HA-DOPAfacilitate liposome lipid binding to various surfaces.

Example 4 In Vivo Effects of Dopamine-Functionalized Hyaluronic Acid andLiposomes

A solution containing 11 mM of HSPC small unilamellar vesicles (SUVs)and 1.6 mg/ml of dopamine-functionalized hyaluronic acid (HA-DOPA;prepared according to procedures described in Example 3) in phosphatebuffer saline (PBS) is injected into animal joints. The level ofdopamine groups in the HA-DOPA is 12% (relative to the number ofrepeating (disaccharide) units of HA). For comparison, correspondingsolutions containing HSPC SUVs and unmodified hyaluronic acid and/orHSPC SUVs without HA are also injected into animal joints.

Retention times of HSPC liposomes injected into joints with HA-DOPA,unmodified HA and/or without HA are compared, by labeling the liposomeswith a fluorescent dye (e.g., IR-783, obtained from Sigma-Aldrich) andmeasuring fluorescent intensity over time.

Therapeutic parameters associated with decreased friction in the jointsare optionally measured in order to evaluate the effect of theadministered solution in vivo.

Although the invention has been described in conjunction with specificembodiments thereof, it is evident that many alternatives, modificationsand variations will be apparent to those skilled in the art.Accordingly, it is intended to embrace all such alternatives,modifications and variations that fall within the spirit and broad scopeof the appended claims.

It is the intent of the applicant(s) that all publications, patents andpatent applications referred to in this specification are to beincorporated in their entirety by reference into the specification, asif each individual publication, patent or patent application wasspecifically and individually noted when referenced that it is to beincorporated herein by reference. In addition, citation oridentification of any reference in this application shall not beconstrued as an admission that such reference is available as prior artto the present invention. To the extent that section headings are used,they should not be construed as necessarily limiting. In addition, anypriority document(s) of this application is/are hereby incorporatedherein by reference in its/their entirety.

What is claimed is:
 1. A method of reducing a friction coefficient of asurface, the method comprising contacting the surface with a solutioncomprising at least one water-soluble polymer, liposomes, and an aqueouscarrier, and modifying the surface so as to obtain a modified surface,wherein said water-soluble polymer and said modified surface areselected such that said water-soluble polymer is attachable to saidmodified surface.
 2. The method of claim 1, wherein a molar percentageof phosphatidylcholine in said liposomes is at least 50%.
 3. The methodof claim 1, wherein said at least one water-soluble polymer comprises anon-ionic polymer.
 4. The method of claim 1, wherein said at least onewater-soluble polymer comprises an ionic polymer.
 5. The method of claim1, wherein said at least one water-soluble polymer comprises abiopolymer.
 6. The method of claim 1, wherein said water-soluble polymeris selected from the group consisting of a hyaluronic acid, apolyvinylpyrrolidone and a polyethylene oxide.
 7. The method of claim 1,wherein said surface is a physiological surface, and said carrier is aphysiologically acceptable carrier.
 8. The method of claim 7, whereinsaid surface is an articular surface of a synovial joint.
 9. The methodof claim 8, being for use in the treatment of a synovial joint disorderassociated with an increased friction coefficient of an articularsurface in the synovial joint.
 10. A method of reducing a frictioncoefficient of a surface, the method comprising: attaching at least onewater-soluble polymer to the surface, wherein said attaching at leastone water-soluble polymer to the surface comprises modifying the surfaceto obtain a modified surface, and said water-soluble polymer is selectedto be attachable to said modified surface; and contacting said at leastone water-soluble polymer with liposomes, thereby effecting coating ofthe surface by an amphiphilic lipid of said liposomes, wherein saidamphiphilic lipid comprises at least one charged group, and wherein atleast a portion of molecules of said amphiphilic lipid coating saidsurface are oriented such that charged groups thereof face outwards atsaid surface.
 11. The method of claim 10, wherein said attaching atleast one water-soluble polymer to the surface is effected prior to saidcontacting said at least one water-soluble polymer with liposomes. 12.The method of claim 10, wherein a molar percentage ofphosphatidylcholine in said liposomes is at least 50%.
 13. The method ofclaim 10, wherein said at least one water-soluble polymer comprises anon-ionic polymer.
 14. The method of claim 10, wherein said at least onewater-soluble polymer comprises an ionic polymer.
 15. The method ofclaim 10, wherein said at least one water-soluble polymer comprises abiopolymer.
 16. The method of claim 10, wherein said water-solublepolymer is selected from the group consisting of a hyaluronic acid, apolyvinylpyrrolidone and a polyethylene oxide.
 17. The method of claim10, wherein said surface is a physiological surface.
 18. The method ofclaim 17, wherein said surface is an articular surface of a synovialjoint.
 19. The method of claim 18, being for use in the treatment of asynovial joint disorder associated with an increased frictioncoefficient of an articular surface in the synovial joint.
 20. Anarticle of manufacture comprising a composition-of-matter, thecomposition-of-matter comprising a substrate coated, on at least aportion of a surface thereof, by at least one water-soluble polymer, thearticle of manufacture being identified for use for efficientlyattaching thereto an amphiphilic lipid so as to reduce a frictioncoefficient of said substrate.